Isoindoline compounds and methods of their use

ABSTRACT

Novel isoindoline compounds are disclosed. Methods of treating, preventing and/or managing cancer, diseases and disorders associated with, or characterized by, undesired angiogenesis, and diseases and disorders mediated by PDE 4, using the compounds are also disclosed.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a divisional application of U.S. patent applicationSer. No. 12/070,322, filed Feb. 15, 2008, now U.S. Pat. No. 7,569,597,which claims the benefit of U.S. patent application Ser. No. 10/900,332,filed Jul. 28, 2004, now U.S. Pat. No. 7,405,237, the disclosures ofwhich are incorporated by reference in their entirety.

1. FIELD OF THE INVENTION

This invention relates to novel isoindoline compounds and their use inmethods of treating, preventing and/or managing cancer, and otherdiseases and disorders including, but not limited to, those associatedwith, or characterized by, undesired angiogenesis and/or those mediatedby PDE 4 inhibition.

2. BACKGROUND OF THE INVENTION

2.1 Pathobiology of Cancer and Other Diseases

Cancer is characterized primarily by an increase in the number ofabnormal cells derived from a given normal tissue, invasion of adjacenttissues by these abnormal cells, or lymphatic or blood-borne spread ofmalignant cells to regional lymph nodes and to distant sites(metastasis). Clinical data and molecular biologic studies indicate thatcancer is a multistep process that begins with minor preneoplasticchanges, which may under certain conditions progress to neoplasia. Theneoplastic lesion may evolve clonally and develop an increasing capacityfor invasion, growth, metastasis, and heterogeneity, especially underconditions in which the neoplastic cells escape the host's immunesurveillance. Roitt, I., Brostoff, J and Kale, D., Immunology,17.1-17.12 (3rd ed., Mosby, St. Louis, Mo., 1993).

There is an enormous variety of cancers which are described in detail inthe medical literature. Examples includes cancer of the lung, colon,rectum, prostate, breast, brain, and intestine. The incidence of cancercontinues to climb as the general population ages, as new cancersdevelop, and as susceptible populations (e.g., people infected with AIDSor excessively exposed to sunlight) grow. A tremendous demand thereforeexists for new methods and compositions that can be used to treatpatients with cancer.

Many types of cancers are associated with new blood vessel formation, aprocess known as angiogenesis. Several of the mechanisms involved intumor-induced angiogenesis have been elucidated. The most direct ofthese mechanisms is the secretion by the tumor cells of cytokines withangiogenic properties. Examples of these cytokines include acidic andbasic fibroblastic growth factor (a,b-FGF), angiogenin, vascularendothelial growth factor (VEGF), and TNF-α Alternatively, tumor cellscan release angiogenic peptides through the production of proteases andthe subsequent breakdown of the extracellular matrix where somecytokines are stored (e.g., b-FGF). Angiogenesis can also be inducedindirectly through the recruitment of inflammatory cells (particularlymacrophages) and their subsequent release of angiogenic cytokines (e.g.,TNF-α, bFGF).

A variety of other diseases and disorders are also associated with, orcharacterized by, undesired angiogenesis. For example, enhanced orunregulated angiogenesis has been implicated in a number of diseases andmedical conditions including, but not limited to, ocular neovasculardiseases, choroidal neovascular diseases, retina neovascular diseases,rubeosis (neovascularization of the angle), viral diseases, geneticdiseases, inflammatory diseases, allergic diseases, and autoimmunediseases. Examples of such diseases and conditions include, but are notlimited to: diabetic retinopathy; retinopathy of prematurity; cornealgraft rejection; neovascular glaucoma; retrolental fibroplasia; andproliferative vitreoretinopathy.

Accordingly, compounds that can control angiogenesis or inhibit theproduction of certain cytokines, including TNF-α, may be useful in thetreatment and prevention of various diseases and conditions.

2.2 Methods of Treatment

Current cancer therapy may involve surgery, chemotherapy, hormonaltherapy and/or radiation treatment to eradicate neoplastic cells in apatient (see, for example, Stockdale, 1998, Medicine, vol. 3, Rubensteinand Federman, eds., Chapter 12, Section IV). Recently, cancer therapycould also involve biological therapy or immunotherapy. All of theseapproaches pose significant drawbacks for the patient. Surgery, forexample, may be contraindicated due to the health of a patient or may beunacceptable to the patient. Additionally, surgery may not completelyremove neoplastic tissue. Radiation therapy is only effective when theneoplastic tissue exhibits a higher sensitivity to radiation than normaltissue. Radiation therapy can also often elicit serious side effects.Hormonal therapy is rarely given as a single agent. Although hormonaltherapy can be effective, it is often used to prevent or delayrecurrence of cancer after other treatments have removed the majority ofcancer cells. Biological therapies and immunotherapies are limited innumber and may produce side effects such as rashes or swellings,flu-like symptoms, including fever, chills and fatigue, digestive tractproblems or allergic reactions.

With respect to chemotherapy, there are a variety of chemotherapeuticagents available for treatment of cancer. A majority of cancerchemotherapeutics act by inhibiting DNA synthesis, either directly, orindirectly by inhibiting the biosynthesis of deoxyribonucleotidetriphosphate precursors, to prevent DNA replication and concomitant celldivision. Gilman et al., Goodman and Gilman's: The Pharmacological Basisof Therapeutics, Tenth Ed. (McGraw Hill, N.Y.).

Despite availability of a variety of chemotherapeutic agents,chemotherapy has many drawbacks. Stockdale, Medicine, vol. 3, Rubensteinand Federman, eds., ch. 12, sect. 10, 1998. Almost all chemotherapeuticagents are toxic, and chemotherapy causes significant, and oftendangerous side effects including severe nausea, bone marrow depression,and immunosuppression. Additionally, even with administration ofcombinations of chemotherapeutic agents, many tumor cells are resistantor develop resistance to the chemotherapeutic agents. In fact, thosecells resistant to the particular chemotherapeutic agents used in thetreatment protocol often prove to be resistant to other drugs, even ifthose agents act by different mechanism from those of the drugs used inthe specific treatment. This phenomenon is referred to as pleiotropicdrug or multidrug resistance. Because of the drug resistance, manycancers prove refractory to standard chemotherapeutic treatmentprotocols.

Other diseases or conditions associated with, or characterized by,undesired angiogenesis are also difficult to treat. However, somecompounds such as protamine, hepain and steroids have been proposed tobe useful in the treatment of certain specific diseases. Taylor et al.,Nature 297:307 (1982); Folkman et al., Science 221:719 (1983); and U.S.Pat. Nos. 5,001,116 and 4,994,443. Thalidomide and certain derivativesof it have also been proposed for the treatment of such diseases andconditions. U.S. Pat. Nos. 5,593,990, 5,629,327, 5,712,291, 6,071,948and 6,114,355 to D'Amato. Additional compounds that are reportedlyeffective are described by U.S. Pat. Nos. 6,380,239 and 6,326,388, bothof which are incorporated in their entirety by reference.

Still, there is a significant need for safe and effective methods oftreating, preventing and managing cancer and other diseases andconditions, particularly for diseases that are refractory to standardtreatments, such as surgery, radiation therapy, chemotherapy andhormonal therapy, while reducing or avoiding the toxicities and/or sideeffects associated with the conventional therapies.

2.3 PDE 4

Adenosine 3′,5′-cyclic monophosphate (cAMP) is another enzyme that playsa role in many diseases and conditions, such as, but not limited toasthma and inflammation (Lowe and Cheng, Drugs of the Future, 17(9),799-807, 1992). The elevation of cAMP in inflammatory leukocytesreportedly inhibits their activation and the subsequent release ofinflammatory mediators, including TNF-α and nuclear factor κB (NF-κB).Increased levels of cAMP also lead to the relaxation of airway smoothmuscle.

It is believed that primary cellular mechanism for the inactivation ofcAMP is the breakdown of cAMP by a family of isoenzymes referred to ascyclic nucleotide phosphodiesterases (PDE) (Beavo and Reitsnyder, Trendsin Pharm., 11, 150-155, 1990). There are twelve known members of thefamily of PDEs. It is recognized that the inhibition of PDE type IV(PDE4) is particularly effective in both the inhibition of inflammatorymediated release and the relaxation of airway smooth muscle (Verghese,et al., Journal of Pharmacology and Experimental Therapeutics, 272(3),1313-1320, 1995). Thus, compounds that specifically inhibit PDE 4 mayinhibit inflammation and aid the relaxation of airway smooth muscle witha minimum of unwanted side effects, such as cardiovascular oranti-platelet effects.

The PDE 4 family that is specific for cAMP is currently the largest, andis composed of at least 4 isozymes (a-d), and multiple splice variants(Houslay, M. D. et al. in Advances in Pharmacology 44, eds. J. August etal., p. 225, 1998). There may be over 20 PDE 4 isoforms expressed in acell specific pattern regulated by a number of different promoters.Disease states for which selective PDE4 inhibitors have been soughtinclude: asthma, atopic dermatitis, depression, reperfusion injury,septic shock, toxic shock, endotoxic shock, adult respiratory distresssyndrome, autoimmune diabetes, diabetes insipidus, multi-infarctdementia, AIDS, cancer, Crohn's disease, multiple sclerosis, cerebralischemia, psoriasis, allograft rejection, restenosis, ulceratiavecolitis, cachexia, cerebral malaria, allergic rhino-conjunctivitis,osteoarthritis, rheumatoid arthritis, chronic obstructive pulmonarydisease (COPD), chronic bronchitis, cosinophilic granuloma, andautoimmune encephalomyelitis (Houslay et al., 1998). PDE 4 is present inthe brain and major inflammatory cells and has been found in abnormallyelevated levels in a number of diseases including atopic dermatitis oreczema, asthma, and hay fever among others (reference OHSU flyer and J.of Allergy and Clinical Immunology, 70: 452-457, 1982 by Grewe et al.).In individuals suffering from atopic diseases elevated PDE 4 activity isfound in their peripheral blood mononuclear leukocytes, T cells, mastcells, neutrophils and basophils. This increased PDE activity decreasescAMP levels and results in a breakdown of cAMP control in these cells.This results in increased immune responses in the blood and tissues ofthose that are affected.

Some PDE 4 inhibitors reportedly have a broad spectrum ofanti-inflammatory activity, with impressive activity in models ofasthma, chronic obstructive pulmonary disorder (COPD) and other allergicdisorders such as atopic dermatitis and hay fever. PDE 4 inhibitors thathave been used include theophylline, rolipram, denbufylline, ARIFLO,ROFLUMILAST, CDP 840 (a tri-aryl ethane) and CP80633 (a pyrimidone). PDE4 inhibitors have been shown to influence eosinophil responses, decreasebasophil histamine release, decrease IgE, PGE2, IL10 synthesis, anddecrease anti-CD3 stimulated Il-4 production. Similarly, PDE4 inhibitorshave been shown to block neutrophil functions. Neutrophils play a majorrole in asthma, chronic obstructive pulmonary disorder (COPD) and otherallergic disorders. PDE 4 inhibitors have been shown to inhibit therelease of adhesion molecules, reactive oxygen species, interleukin(IL)-8 and neutrophil elastase, associated with neutrophils whichdisrupt the architecture of the lung and therefore airway function. PDE4 inhibitors influence multiple functional pathways, act on multipleimmune and inflammatory pathways, and influence synthesis or release ofnumerous immune mediators. J. M. Hanifin and S. C. Chan, “AtopicDermatitis-Therapeutic Implication for New PhosphodiesteraseInhibitors,” Monocyte Dysregulation of T Cells in AACI News, 7/2, 1995;J. M. Hanifin et al., “Type 4 Phosphodiesterase Inhibitors Have clinicaland In Vitro Anti-inflammatory Effects in Atopic Dermatitis,” Journal ofInvestigative Dermatology, 1996, 107, pp 51-56).

Some of the first generation of PDE 4 inhibitors are effective ininhibiting PDE4 activity and alleviating a number of the inflammatoryproblems caused by over expression of this enzyme. However, theireffectiveness is limited by side effects, particularly when usedsystemically, such as nausea and vomiting. Huang et al., Curr. Opin. InChem. Biol. 2001, 5:432-438. Indeed, many of the PDE 4 inhibitorsdeveloped to date have been small molecule compounds with centralnervous system and gastrointestinal side effects, e.g., headache,nausea/emesis, and gastric secretion.

3. SUMMARY OF THE INVENTION

This invention encompasses novel isoindoline compounds andpharmaceutically acceptable salts, solvates, prodrugs, and stereoisomersthereof.

This invention also encompasses methods of treating and preventingcancer. This invention also encompasses methods of treating andpreventing certain types of cancer, including primary and metastaticcancer, as well as cancers that are refractory or resistant toconventional chemotherapy. The methods comprise administering to apatient in need of such treatment or prevention a therapeutically orprophylactically effective amount of a compound of this invention, or apharmaceutically acceptable salt, solvate, stereoisomer, or prodrugthereof.

The invention also encompasses methods of managing cancer (e.g.,preventing or prolonging their recurrence, or lengthening the time ofremission), which comprise administering to a patient in need of suchmanagement a prophylactically effective amount of a compound of thisinvention, or a pharmaceutically acceptable salt, solvate, hydrate,stereoisomer, clathrate, or prodrug thereof.

In particular methods of the invention, a compound of the invention isadministered in combination with a therapy conventionally used to treat,prevent or manage cancer. Examples of such conventional therapiesinclude, but are not limited to, surgery, chemotherapy, radiationtherapy, hormonal therapy, biological therapy and immunotherapy.

This invention also encompasses methods of treating, managing andpreventing diseases and disorders other than cancer that are associatedwith, or characterized by, undesired angiogenesis, which compriseadministering to a patient in need of such treatment, management orprevention a therapeutically or prophylactically effective amount of acompound of this invention, or a pharmaceutically acceptable salt,solvate, hydrate, stereoisomer, clathrate, or prodrug thereof.

This invention also encompasses methods of treating, managing andpreventing diseases and disorders mediated by PDE 4, which compriseadministering to a patient in need of such treatment, management orprevention a therapeutically or prophylactically effective amount of acompound of this invention, or a pharmaceutically acceptable salt,solvate, hydrate, stereoisomer, clathrate, or prodrug thereof.

In other methods of the invention, a compound of the invention isadministered in combination with a therapy conventionally used to treat,prevent or manage diseases or disorders associated with, orcharacterized by, undesired angiogenesis or mediated by PDE 4. Examplesof such conventional therapies include, but are not limited to, surgery,chemotherapy, radiation therapy, hormonal therapy, biological therapyand immunotherapy.

This invention encompasses pharmaceutical compositions, single unitdosage forms, dosing regimens and kits which comprise a compound of thisinvention, and optionally a second, or additional, active agent. Secondactive agents include specific combinations, or “cocktails,” of drugs.

4. DETAILED DESCRIPTION OF THE INVENTION

This invention encompasses novel isoindoline compounds described herein,and pharmaceutically acceptable salts, solvates, prodrugs, andstereoisomers thereof.

Another embodiment of the invention encompasses methods of treating,managing, or preventing cancer which comprises administering to apatient in need of such treatment or prevention a therapeutically orprophylactically effective amount of a compound of the invention, or apharmaceutically acceptable salt, solvate, hydrate, stereoisomer,clathrate, or prodrug thereof.

In particular methods encompassed by this embodiment, a compound of thisinvention is administered in combination with another pharmacologicallyactive agent (“second active agent”) or in conjunction with anothermethod of treating, managing, or preventing cancer. Methods, ortherapies, that can be used in combination with the administration of acompound of this invention include, but are not limited to, surgery,blood transfusions, immunotherapy, biological therapy, radiationtherapy, and other non-drug based therapies presently used to treat,prevent or manage cancer.

Another embodiment of the invention encompasses methods of treating,managing or preventing diseases and disorders other than cancer that arecharacterized by undesired angiogenesis. These methods comprise theadministration of a therapeutically or prophylactically effective amountof a compound of the invention.

Examples of diseases and disorders associated with, or characterized by,undesired angiogenesis include, but are not limited to, inflammatorydiseases, autoimmune diseases, viral diseases, genetic diseases,allergic diseases, bacterial diseases, ocular neovascular diseases,choroidal neovascular diseases, retina neovascular diseases, andrubeosis (neovascularization of the angle). Specific examples of thediseases and disorders associated with, or characterized by, undesiredangiogenesis include, but are not limited to, endometriosis, Crohn'sdisease, heart failure, advanced heart failure, renal impairment,endotoxemia, toxic shock syndrome, osteoarthritis, retrovirusreplication, wasting, meningitis, silica-induced fibrosis,asbestos-induced fibrosis, veterinary disorder, malignancy-associatedhypercalcemia, stroke, circulatory shock, periodontitis, gingivitis,macrocytic anemia, refractory anemia, and 5q-syndrome.

Another embodiment of this invention encompasses methods of inhibitingor reducing PDE 4 comprising contacting a compound of this inventionwith PDE 4. This invention also encompasses methods of inhibiting orreducing the level or activity of PDE 4 in a patient (e.g., human)comprising administering a compound of this invention to the patient.This invention also encompasses methods of treating, managing orpreventing diseases and disorders mediated by PDE 4, which comprise theadministration of a therapeutically or prophylactically effective amountof a compound of the invention.

Examples of diseases or disorders mediated by PDE 4 include, but are notlimited to, asthma, atopic dermatitis, depression, reperfusion injury,septic shock, toxic shock, endotoxic shock, adult respiratory distresssyndrome, autoimmune diabetes, diabetes insipidus, multi-infarctdementia, AIDS, cancer, Crohn's disease, multiple sclerosis, cerebralischemia, psoriasis, allograft rejection, restenosis, ulceratiavecolitis, cachexia, cerebral malaria, allergic rhino-conjunctivitis,osteoarthritis, rheumatoid arthritis, chronic obstructive pulmonarydisease (COPD), chronic bronchitis, cosinophilic granuloma, andautoimmune encephalomyelitis.

In particular methods encompassed by this embodiment, a compound of theinvention is administered in combination with a second active agent, orin addition to other methods of treating, managing, or preventing thedisease or condition. Second active agents include small molecules andlarge molecules (e.g., proteins and antibodies), examples of which areprovided herein, as well as stem cells. Methods, or therapies, that canbe used in combination with the administration of a compound of thisinvention include, but are not limited to, surgery, blood transfusions,immunotherapy, biological therapy, radiation therapy, and other non-drugbased therapies presently used to treat, prevent or manage disease andconditions associated with, or characterized by, undesired angiogenesis.

The invention also encompasses pharmaceutical compositions (e.g., singleunit dosage forms) that can be used in methods disclosed herein.Particular pharmaceutical compositions comprise a compound of theinvention and optionally a second active agent.

4.1 Compounds of the Invention

This invention encompasses novel isoindoline compounds, andpharmaceutically acceptable salts, solvates, prodrugs, and stereoisomersthereof, which are described herein.

In one embodiment, this invention encompasses compounds of formula I,and pharmaceutically acceptable salts, solvates, prodrugs, andstereoisomers thereof:

in which the carbon atom designated C* constitutes a center of chirality(when n is not zero and R¹ is not the same as R²); one of X¹ and X² isamino, nitro, alkyl of one to six carbons, or NH—Z, and the other of X¹or X² is hydrogen; each of R¹ and R² independent of the other, ishydroxy or NH—Z; R³ is hydrogen, alkyl of one to six carbons, halo, orhaloalkyl; Z is hydrogen, aryl, alkyl of one to six carbons, formyl, oracyl of one to six carbons; and n has a value of 0, 1, or 2; providedthat if X¹ is amino, and n is 1 or 2, then R¹ and R² are not bothhydroxy.

In another embodiment, this invention encompasses compounds of formulaII, and pharmaceutically acceptable salts, solvates, prodrugs, andstereoisomers thereof:

in which the carbon atom designated C* constitutes a center of chiralitywhen n is not zero and R¹ is not R²; one of X¹ and X² is amino, nitro,alkyl of one to six carbons, or NH—Z, and the other of X¹ or X² ishydrogen; each of R¹ and R² independent of the other, is hydroxy orNH—Z; R³ is alkyl of one to six carbons, halo, or hydrogen; Z ishydrogen, aryl or an alkyl or acyl of one to six carbons; and n has avalue of 0, 1, or 2.

Specific examples include, but are not limited to,2-(4-amino-1-oxo-1,3-dihydro-isoindol-2-yl)-4-carbamoyl-butyric acid and4-(4-amino-1-oxo-1,3-dihydro-isoindol-2-yl)-4-cabamoyl-butyric acid,which have the following structures, respectively, and pharmaceuticallyacceptable salts, solvates, prodrugs, and stereoisomers thereof:

Other representative compounds are of formula:

in which the carbon atom designated C* constitutes a center of chiralitywhen n is not zero and R¹ is not R²; one of X¹ and X² is amino, nitro,alkyl of one to six carbons, or NH—Z, and the other of X¹ or X² ishydrogen; each of R¹ and R² independent of the other, is hydroxy orNH—Z; R³ is alkyl of one to six carbons, halo, or hydrogen; Z ishydrogen, aryl, an alkyl or acyl of one to six carbons, or furanylalkyl,wherein the alkyl has one to six carbons; and n has a value of 0, 1, or2; and the pharmaceutically acceptable salts, solvates, prodrugs, andstereoisomers thereof.

Specific examples include, but are not limited to,4-carbamoyl-4-{4-[(furan-2-yl-methyl)-amino]-1,3-dioxo-1,3-dihydro-isoindol-2-yl}-butyricacid,4-carbamoyl-2-{4-[(furan-2-yl-methyl)-amino]-1,3-dioxo-1,3-dihydro-isoindol-2-yl}-butyricacid,2-{4-[(furan-2-yl-methyl)-amino]-1,3-dioxo-1,3-dihydro-isoindol-2-yl}-4-phenylcarbamoyl-butyricacid, and2-{4-[(furan-2-yl-methyl)-amino]-1,3-dioxo-1,3-dihydro-isoindol-2-yl}-pentanedioicacid, which have the following structures, respectively, andpharmaceutically acceptable salts, solvate, prodrugs, and stereoisomersthereof:

Other specific examples of the compounds are of formula:

wherein one of X¹ and X² is nitro, or NH-Z, and the other of X¹ or X² ishydrogen;

each of R¹ and R², independent of the other, is hydroxy or NH—Z;

R³ is alkyl of one to six carbons, halo, or hydrogen;

Z is hydrogen, phenyl, an acyl of one to six carbons, or an alkyl of oneto six carbons; and

n has a value of 0, 1, or 2;

provided that if one of X¹ and X² is nitro, and n is 1 or 2, then R¹ andR² are other than hydroxy; and

if —COR¹ and —(CH₂)_(n)COR² are different, the carbon atom designated C*constitutes a center of chirality. Other representative compounds are offormula:

wherein one of X¹ and X² is alkyl of one to six carbons;

each of R¹ and R², independent of the other, is hydroxy or NH—Z;

R³ is alkyl of one to six carbons, halo, or hydrogen;

Z is hydrogen, phenyl, an acyl of one to six carbons, or an alkyl of oneto six carbons; and

n has a value of 0, 1, or 2; and

if —COR¹ and —(CH₂)_(n)COR² are different, the carbon atom designated C*constitutes a center of chirality.

In another embodiment, this invention encompasses compounds of formulaIII, and pharmaceutically acceptable salts, solvates, prodrugs, andstereoisomers thereof:

wherein:

the carbon atom designated* constitutes a center of chirality;

Y is C═O, CH₂, SO₂ or CH₂C═O;

X is hydrogen, or alkyl of 1 to 4 carbon atoms;

(i) each of R¹, R², R³, and R⁴, independently of the others, ishydrogen, halo, trifluoromethyl, acetyl, alkyl of 1 to 8 carbon atoms,alkoxy of 1 to 4 carbon atoms, nitro, cyano, hydroxy, —CH₂NR⁸R⁹,—(CH₂)₂NR⁸R⁹, or —NR⁸R⁹ or

(ii) any two of R¹, R², R³, and R⁴ on adjacent carbon atoms, togetherwith the depicted benzene ring to which they are bound arenaphthylidene, quinoline, quinoxaline, benzimidazole, benzodioxole or2-hydroxybenzimidazole;

each of R⁵ and R⁶, independently of the other, is hydrogen, alkyl of 1to 4 carbon atoms, alkoxy of 1 to 6 carbon atoms, cyano,benzocycloalkoxy, cycloalkoxy of up to 18 carbon atoms, bicyloalkoxy ofup to 18 carbon atoms, tricylcoalkoxy of up to 18 carbon atoms, orcycloalkylalkoxy of up to 18 carbon atoms;

(i) each of R⁸ and R⁹, independently of the other, is hydrogen, alkyl of1 to 8 carbon atoms, phenyl, benzyl, pyridyl, pyridylmethyl, or

(ii) one of R⁸ and R⁹ is hydrogen and the other is —COR¹⁰, or —SO₂R¹⁰,in which R¹⁰ is hydrogen, alkyl of 1 to 8 carbon atoms, cycloalkyl,cycloalkylmethyl of up to 6 carbon atoms, phenyl, pyridyl, benzyl,imidazolylmethyl, pyridylmethyl, NR¹¹R¹², or CH₂NR¹⁴R¹⁵, wherein R¹¹ andR¹², independently of each other, are hydrogen, alkyl of 1 to 8 carbonatoms, phenyl, or benzyl and R¹⁴ and R¹⁵, independently of each other,are hydrogen, methyl, ethyl, or propyl; or

(iii) R⁸ and R⁹ taken together are tetramethylene, pentamethylene,hexamethylene, —CH═NCH═CH—, or —CH₂CH₂X¹CH₂CH₂— in which X¹ is —O—, —S—,or —NH—.

Specific examples include, but are not limited to,cyclopropanecarboxylic acid{2-[1-(3-ethoxy-4-methoxy-phenyl)-2-[1,3,4]oxadiazol-2-yl-ethyl]-3-oxo-2,3-dihydro-1H-isoindol-4-yl}-amide,which has the following chemical structure, and pharmaceuticallyacceptable salts, solvates, prodrugs, and stereoisomers thereof:

As used herein and unless otherwise indicated, the term“pharmaceutically acceptable salt” encompasses non-toxic acid and baseaddition salts of the compound to which the term refers. Acceptablenon-toxic acid addition salts include those derived from organic andinorganic acids or bases known in the art, which include, for example,hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid,methanesulphonic acid, acetic acid, tartaric acid, lactic acid, succinicacid, citric acid, malic acid, maleic acid, sorbic acid, aconitic acid,salicylic acid, phthalic acid, embolic acid, enanthic acid, and thelike.

Compounds that are acidic in nature are capable of forming salts withvarious pharmaceutically acceptable bases. The bases that can be used toprepare pharmaceutically acceptable base addition salts of such acidiccompounds are those that form non-toxic base addition salts, i.e., saltscontaining pharmacologically acceptable cations such as, but not limitedto, alkali metal or alkaline earth metal salts and the calcium,magnesium, sodium or potassium salts in particular. Suitable organicbases include, but are not limited to, N,N-dibenzylethylenediamine,chloroprocaine, choline, diethanolamine, ethylenediamine, meglumaine(N-methylglucamine), lysine, and procaine.

Neutral forms of some compounds may be regenerated by contacting thesalt with a base or acid and isolating the parent compound in theconventional manner. The parent form of a compound can differ from itsvarious salt forms in certain physical properties, such as solubility inpolar solvents, but the salts are typically equivalent to the parentform of the compound for the purposes of the present invention.

As used herein and unless otherwise indicated, the term “prodrug” meansa derivative of a compound that can hydrolyze, oxidize, or otherwisereact under biological conditions (in vitro or in vivo) to provide thecompound. Examples of prodrugs include, but are not limited to,derivatives of compounds of this invention that comprise biohydrolyzablemoieties such as biohydrolyzable amides, biohydrolyzable esters,biohydrolyzable carbamates, biohydrolyzable carbonates, biohydrolyzableureides, and biohydrolyzable phosphate analogues. Other examples ofprodrugs include derivatives of a compound of this invention thatcomprise —NO, —NO₂, —ONO, or —ONO₂ moieties. Prodrugs can typically beprepared using well-known methods, such as those described in 1 Burger'sMedicinal Chemistry and Drug Discovery, 172-178, 949-982 (Manfred E.Wolff ed., 5th ed. 1995), and Design of Prodrugs (H. Bundgaard ed.,Elselvier, N.Y. 1985).

As used herein and unless otherwise indicated, the terms“biohydrolyzable amide,” “biohydrolyzable ester,” “biohydrolyzablecarbamate,” “biohydrolyzable carbonate,” “biohydrolyzable ureide,” and“biohydrolyzable phosphate” mean an amide, ester, carbamate, carbonate,ureide, or phosphate, respectively, of a compound that either: 1) doesnot interfere with the biological activity of the compound but canconfer upon that compound advantageous properties in vivo, such asuptake, duration of action, or onset of action; or 2) is biologicallyinactive but is converted in vivo to the biologically active compound.Examples of biohydrolyzable esters include, but are not limited to,lower alkyl esters, lower acyloxyalkyl esters (such as acetoxylmethyl,acetoxyethyl, aminocarbonyloxymethyl, pivaloyloxymethyl, andpivaloyloxyethyl esters), lactonyl esters (such as phthalidyl andthiophthalidyl esters), lower alkoxyacyloxyalkyl esters (such asmethoxycarbonyloxymethyl, ethoxycarbonyloxyethyl andisopropoxycarbonyloxyethyl esters), alkoxyalkyl esters, choline esters,and acylamino alkyl esters (such as acetamidomethyl esters). Examples ofbiohydrolyzable amides include, but are not limited to, lower alkylamides, α-amino acid amides, alkoxyacyl amides, andalkylaminoalkylcarbonyl amides. Examples of biohydrolyzable carbamatesinclude, but are not limited to, lower alkylamines, substitutedethylenediamines, aminoacids, hydroxyalkylamines, heterocyclic andheteroaromatic amines, and polyether amines.

Various compounds of the invention contain one or more chiral centers,and can exist as racemic mixtures of enantiomers or mixtures ofdiastereomers. This invention encompasses the use of stereomericallypure forms of such compounds, as well as the use of mixtures of thoseforms. For example, mixtures comprising equal or unequal amounts of theenantiomers of compounds of this invention may be used in methods andcompositions of the invention. The purified (R) or (S) enantiomers ofthe specific compounds disclosed herein may be used substantially freeof its other enantiomer.

As used herein and unless otherwise indicated, the term “stereomericallypure” means a composition that comprises one stereoisomer of a compoundand is substantially free of other stereoisomers of that compound. Forexample, a stereomerically pure composition of a compound having onechiral center will be substantially free of the opposite enantiomer ofthe compound. A stereomerically pure composition of a compound havingtwo chiral centers will be substantially free of other diastereomers ofthe compound. A typical stereomerically pure compound comprises greaterthan about 80% by weight of one stereoisomer of the compound and lessthan about 20% by weight of other stereoisomers of the compound, morepreferably greater than about 90% by weight of one stereoisomer of thecompound and less than about 10% by weight of the other stereoisomers ofthe compound, even more preferably greater than about 95% by weight ofone stereoisomer of the compound and less than about 5% by weight of theother stereoisomers of the compound, and most preferably greater thanabout 97% by weight of one stereoisomer of the compound and less thanabout 3% by weight of the other stereoisomers of the compound.

As used herein and unless otherwise indicated, the term “stereomericallyenriched” means a composition that comprises greater than about 60% byweight of one stereoisomer of a compound, preferably greater than about70% by weight, more preferably greater than about 80% by weight of onestereoisomer of a compound.

As used herein and unless otherwise indicated, the term“enantiomerically pure” means a stereomerically pure composition of acompound having one chiral center. Similarly, the term “enantiomericallyenriched” means a stereomerically enriched composition of a compoundhaving one chiral center.

Certain compounds of the present invention can exist in unsolvated formsas well as solvated forms, including hydrated forms. In general,solvated forms are equivalent to unsolvated forms. Certain compounds ofthe invention may exist in multiple crystalline or amorphous forms. Ingeneral, all physical forms are equivalent for the uses contemplated bythe present invention, and are encompassed by the present invention.

Certain compounds of the present invention possess asymmetric carbonatoms (optical or stereo-centers) or double bonds; the racemates,enantiomers, diastereomers, geometric isomers and mixtures thereof areall intended to be encompassed by this invention.

Compounds of the invention may also contain unnatural proportions ofatomic isotopes at one or more of the atoms that constitute suchcompounds. For example, the compounds may be radiolabeled withradioactive isotopes, such as for example tritium (³H), iodine-125(¹²⁵I) or carbon-14 (¹⁴C). Radiolabeled compounds are useful astherapeutic agents, e.g., cancer therapeutic agents, research reagents,e.g., assay reagents, and diagnostic agents, e.g., in vivo imagingagents. All isotopic variations of the compounds of the presentinvention, whether radioactive or not, are intended to be encompassedwithin the scope of the present invention.

It should be noted that if there is a discrepancy between a depictedstructure and a name given that structure, the depicted structure is tobe accorded more weight. In addition, if the stereochemistry of astructure or a portion of a structure is not indicated with, forexample, bold or dashed lines, the structure or portion of the structureis to be interpreted as encompassing all stereoisomers of it.

4.2 Second Active Agents

Compounds of this invention can be combined with other pharmacologicallyactive compounds (“second active agents”) in methods and compositions ofthe invention. It is believed that certain combinations worksynergistically in the treatment of particular types of cancer andcertain diseases and conditions associated with, or characterized by,undesired angiogenesis. Compounds of this invention can also work toalleviate adverse effects associated with certain second active agents,and some second active agents can be used to alleviate adverse effectsthat may be associated with compounds of this invention.

One or more second active ingredients or agents can be used in themethods and compositions of the invention together with a compound ofthe invention. Second active agents can be large molecules (e.g.,proteins) or small molecules (e.g., synthetic inorganic, organometallic,or organic molecules).

Examples of large molecule active agents include, but are not limitedto, hematopoietic growth factors, cytokines, and monoclonal andpolyclonal antibodies. Specific examples of the active agents areanti-CD40 monoclonal antibodies (such as, for example, SGN-40); histonedeacetylyase inhibitors (such as, for example, SAHA and LAQ 824);heat-shock protein-90 inhibitors (such as, for example, 17-AAG);insulin-like growth factor-1 receptor kinase inhibitors; vascularendothelial growth factor receptor kinase inhibitors (such as, forexample, PTK787); insulin growth factor receptor inhibitors;lysophosphatidic acid acyltransrerase inhibitors; IkB kinase inhibitors;p38MAPK inhibitors; EGFR inhibitors (such as, for example, gefitinib anderlotinib HCL); HER-2 antibodies (such as, for example, trastuzumab(Herceptin®) and pertuzumab (Omnitarg™)); VEGFR antibodies (such as, forexample, bevacizumab (Avastin™)); VEGFR inhibitors (such as, forexample, flk-1 specific kinase inhibitors, SU5416 and ptk787/zk222584);P13K inhibitors (such as, for example, wortmannin); C-Met inhibitors(such as, for example, PHA-665752); monoclonal antibodies (such as, forexample, rituximab (Rituxan®), tositumomab (Bexxar®), edrecolomab(Panorex®) and G250); and anti-TNF-α antibodies.

Typical large molecule active agents are biological molecules, such asnaturally occurring or artificially made proteins. Proteins that areparticularly useful in this invention include proteins that stimulatethe survival and/or proliferation of hematopoietic precursor cells andimmunologically active poietic cells in vitro or in vivo. Othersstimulate the division and differentiation of committed erythroidprogenitors in cells in vitro or in vivo. Particular proteins include,but are not limited to: interleukins, such as IL-2 (includingrecombinant IL-II (“rIL2”) and canarypox IL-2), IL-10, IL-12, and IL-18;interferons, such as interferon alfa-2a, interferon alfa-2b, interferonalfa-n1, interferon alfa-n3, interferon beta-I a, and interferon gamma-Ib; GM-CF and GM-CSF; and EPO.

Particular proteins that can be used in the methods and compositions ofthe invention include, but are not limited to: filgrastim, which is soldin the United States under the trade name Neupogen® (Amgen, ThousandOaks, Calif.); sargramostim, which is sold in the United States underthe trade name Leukine® (Immunex, Seattle, Wash.); and recombinant EPO,which is sold in the United States under the trade name Epogen® (Amgen,Thousand Oaks, Calif.).

Recombinant and mutated forms of GM-CSF can be prepared as described inU.S. Pat. Nos. 5,391,485; 5,393,870; and 5,229,496; all of which areincorporated herein by reference. Recombinant and mutated forms of G-CSFcan be prepared as described in U.S. Pat. Nos. 4,810,643; 4,999,291;5,528,823; and 5,580,755; all of which are incorporated herein byreference.

This invention encompasses the use of native, naturally occurring, andrecombinant proteins. The invention further encompasses mutants andderivatives (e.g., modified forms) of naturally occurring proteins thatexhibit, in vivo, at least some of the pharmacological activity of theproteins upon which they are based. Examples of mutants include, but arenot limited to, proteins that have one or more amino acid residues thatdiffer from the corresponding residues in the naturally occurring formsof the proteins. Also encompassed by the term “mutants” are proteinsthat lack carbohydrate moieties normally present in their naturallyoccurring forms (e.g., nonglycosylated forms). Examples of derivativesinclude, but are not limited to, pegylated derivatives and fusionproteins, such as proteins formed by fusing IgG1 or IgG3 to the proteinor active portion of the protein of interest. See, e.g., Penichet, M. L.and Morrison, S. L., J. Immunol. Methods 248:91-101 (2001).

Large molecule active agents may be administered in the form ofanti-cancer vaccines. For example, vaccines that secrete, or cause thesecretion of, cytokines such as IL-2, G-CSF, and GM-CSF can be used inthe methods, pharmaceutical compositions, and kits of the invention.See, e.g., Emens, L. A., et al., Curr. Opinion Mol. Ther. 3(1):77-84(2001).

In one embodiment of the invention, the large molecule active agentreduces, eliminates, or prevents an adverse effect associated with theadministration of a compound of this invention. Depending on theparticular compound and the disease or disorder being treated, adverseeffects may include, but are not limited to, drowsiness and somnolence,dizziness and orthostatic hypotension, neutropenia, infections thatresult from neutropenia, increased HIV-viral load, bradycardia,Stevens-Johnson Syndrome and toxic epidermal necrolysis, and seizures(e.g., grand mal convulsions). A specific adverse effect is neutropenia.

Second active agents that are small molecules can also be used toalleviate adverse effects associated with the administration of acompound of this invention. However, like some large molecules, many arebelieved to be capable of providing a synergistic effect whenadministered with (e.g., before, after or simultaneously) a compound ofthe invention. Examples of small molecule second active agents include,but are not limited to, anti-cancer agents, antibiotics,immunosuppressive agents, and steroids.

Examples of anti-cancer agents include, but are not limited to:semaxanib; cyclosporin; etanercept; doxycycline; bortezomib; acivicin;aclarubicin; acodazole hydrochloride; acronine; adozelesin; aldesleukin;altretamine; ambomycin; ametantrone acetate; amsacrine; anastrozole;anthramycin; asparaginase; asperlin; azacitidine; azetepa; azotomycin;batimastat; benzodepa; bicalutamide; bisantrene hydrochloride; bisnafidedimesylate; bizelesin; bleomycin sulfate; brequinar sodium; bropirimine;busulfan; cactinomycin; calusterone; caracemide; carbetimer;carboplatin; carmustine; carubicin hydrochloride; carzelesin;cedefingol; celecoxib (COX-2 inhibitor); chlorambucil; cirolemycin;cisplatin; cladribine; crisnatol mesylate; cyclophosphamide; cytarabine;dacarbazine; dactinomycin; daunorubicin hydrochloride; decitabine;dexormaplatin; dezaguanine; dezaguanine mesylate; diaziquone; docetaxel;doxorubicin; doxorubicin hydrochloride; droloxifene; droloxifenecitrate; dromostanolone propionate; duazomycin; edatrexate; eflomithinehydrochloride; elsamitrucin; enloplatin; enpromate; epipropidine;epirubicin hydrochloride; erbulozole; esorubicin hydrochloride;estramustine; estramustine phosphate sodium; etanidazole; etoposide;etoposide phosphate; etoprine; fadrozole hydrochloride; fazarabine;fenretinide; floxuridine; fludarabine phosphate; fluorouracil;fluorocitabine; fosquidone; fostriecin sodium; gemcitabine; gemcitabinehydrochloride; hydroxyurea; idarubicin hydrochloride; ifosfamide;ilmofosine; iproplatin; irinotecan; irinotecan hydrochloride; lanreotideacetate; letrozole; leuprolide acetate; liarozole hydrochloride;lometrexol sodium; lomustine; losoxantrone hydrochloride; masoprocol;maytansine; mechlorethamine hydrochloride; megestrol acetate;melengestrol acetate; melphalan; menogaril; mercaptopurine;methotrexate; methotrexate sodium; metoprine; meturedepa; mitindomide;mitocarcin; mitocromin; mitogillin; mitomalcin; mitomycin; mitosper;mitotane; mitoxantrone hydrochloride; mycophenolic acid; nocodazole;nogalamycin; ormaplatin; oxisuran; paclitaxel; pegaspargase; peliomycin;pentamustine; peplomycin sulfate; perfosfamide; pipobroman; piposulfan;piroxantrone hydrochloride; plicamycin; plomestane; porfimer sodium;porfiromycin; prednimustine; procarbazine hydrochloride; puromycin;puromycin hydrochloride; pyrazofurin; riboprine; safingol; safingolhydrochloride; semustine; simtrazene; sparfosate sodium; sparsomycin;spirogermanium hydrochloride; spiromustine; spiroplatin; streptonigrin;streptozocin; sulofenur; talisomycin; tecogalan sodium; taxotere;tegafur; teloxantrone hydrochloride; temoporfin; teniposide; teroxirone;testolactone; thiamiprine; thioguanine; thiotepa; tiazofurin;tirapazamine; toremifene citrate; trestolone acetate; triciribinephosphate; trimetrexate; trimetrexate glucuronate; triptorelin;tubulozole hydrochloride; uracil mustard; uredepa; vapreotide;verteporfin; vinblastine sulfate; vincristine sulfate; vindesine;vindesine sulfate; vinepidine sulfate; vinglycinate sulfate;vinleurosine sulfate; vinorelbine tartrate; vinrosidine sulfate;vinzolidine sulfate; vorozole; zeniplatin; zinostatin; and zorubicinhydrochloride.

Other anti-cancer drugs include, but are not limited to: 20-epi-1,25dihydroxyvitamin D3; 5-ethynyluracil; abiraterone; aclarubicin;acylfulvene; adecypenol; adozelesin; aldesleukin; ALL-TK antagonists;altretamine; ambamustine; amidox; amifostine; aminolevulinic acid;amrubicin; amsacrine; anagrelide; anastrozole; andrographolide;angiogenesis inhibitors; antagonist D; antagonist G; antarelix;anti-dorsalizing morphogenetic protein-1; antiandrogen, prostaticcarcinoma; antiestrogen; antineoplaston; antisense oligonucleotides;aphidicolin glycinate; apoptosis gene modulators; apoptosis regulators;apurinic acid; ara-CDP-DL-PTBA; arginine deaminase; asulacrine;atamestane; atrimustine; axinastatin 1; axinastatin 2; axinastatin 3;azasetron; azatoxin; azatyrosine; baccatin III derivatives; balanol;batimastat; BCR/ABL antagonists; benzochlorins; benzoylstaurosporine;beta lactam derivatives; beta-alethine; betaclamycin B; betulinic acid;bFGF inhibitor; bicalutamide; bisantrene; bisaziridinyispermine;bisnafide; bistratene A; bizelesin; breflate; bropirimine; budotitane;buthionine sulfoximine; calcipotriol; calphostin C; camptothecinderivatives; capecitabine; carboxamide-amino-triazole;carboxyamidotriazole; CaRest M3; CARN 700; cartilage derived inhibitor;carzelesin; casein kinase inhibitors (ICOS); castanospermine; cecropinB; cetrorelix; chlorlns; chloroquinoxaline sulfonamide; cicaprost;cis-porphyrin; cladribine; clomifene analogues; clotrimazole;collismycin A; collismycin B; combretastatin A4; combretastatinanalogue; conagenin; crambescidin 816; crisnatol; cryptophycin 8;cryptophycin A derivatives; curacin A; cyclopentanthraquinones;cycloplatam; cypemycin; cytarabine ocfosfate; cytolytic factor;cytostatin; dacliximab; decitabine; dehydrodidemnin B; deslorelin;dexamethasone; dexifosfamide; dexrazoxane; dexverapamil; diaziquone;didemnin B; didox; diethylnorspermine; dihydro-5-azacytidine;dihydrotaxol, 9-; dioxamycin; diphenyl spiromustine; docetaxel;docosanol; dolasetron; doxifluridine; doxorubicin; droloxifene;dronabinol; duocarmycin SA; ebselen; ecomustine; edelfosine;edrecolomab; eflomithine; elemene; emitefur; epirubicin; epristeride;estramustine analogue; estrogen agonists; estrogen antagonists;etanidazole; etoposide phosphate; exemestane; fadrozole; fazarabine;fenretinide; filgrastim; finasteride; flavopiridol; flezelastine;fluasterone; fludarabine; fluorodaunorunicin hydrochloride; forfenimex;formestane; fostriecin; fotemustine; gadolinium texaphyrin; galliumnitrate; galocitabine; ganirelix; gelatinase inhibitors; gemcitabine;glutathione inhibitors; hepsulfam; heregulin; hexamethylenebisacetamide; hypericin; ibandronic acid; idarubicin; idoxifene;idramantone; ilmofosine; ilomastat; imatinib (e.g., Gleevec®),imiquimod; immunostimulant peptides; insulin-like growth factor-1receptor inhibitor; interferon agonists; interferons; interleukins;iobenguane; iododoxorubicin; ipomeanol, 4-; iroplact; irsogladine;isobengazole; isohomohalicondrin B; itasetron; jasplakinolide;kahalalide F; lamellarin-N triacetate; lanreotide; leinamycin;lenograstim; lentinan sulfate; leptolstatin; letrozole; leukemiainhibiting factor; leukocyte alpha interferon;leuprolide+estrogen+progesterone; leuprorelin; levamisole; liarozole;linear polyamine analogue; lipophilic disaccharide peptide; lipophilicplatinum compounds; lissoclinamide 7; lobaplatin; lombricine;lometrexol; lonidamine; losoxantrone; loxoribine; lurtotecan; lutetiumtexaphyrin; lysofylline; lytic peptides; maitansine; mannostatin A;marimastat; masoprocol; maspin; matrilysin inhibitors; matrixmetalloproteinase inhibitors; menogaril; merbarone; meterelin;methioninase; metoclopramide; MIF inhibitor; mifepristone; miltefosine;mirimostim; mitoguazone; mitolactol; mitomycin analogues; mitonafide;mitotoxin fibroblast growth factor-saporin; mitoxantrone; mofarotene;molgramostim; Erbitux, human chorionic gonadotrophin; monophosphoryllipid A+myobacterium cell wall sk; mopidamol; mustard anticancer agent;mycaperoxide B; mycobacterial cell wall extract; myriaporone;N-acetyldinaline; N-substituted benzamides; nafarelin; nagrestip;naloxone+pentazocine; napavin; naphterpin; nartograstim; nedaplatin;nemorubicin; neridronic acid; nilutamide; nisamycin; nitric oxidemodulators; nitroxide antioxidant; nitrullyn; oblimersen (Genasense®);O⁶-benzylguanine; octreotide; okicenone; oligonucleotides; onapristone;ondansetron; ondansetron; oracin; oral cytokine inducer; ormaplatin;osaterone; oxaliplatin; oxaunomycin; paclitaxel; paclitaxel analogues;paclitaxel derivatives; palauamine; palmitoylrhizoxin; pamidronic acid;panaxytriol; panomifene; parabactin; pazelliptine; pegaspargase;peldesine; pentosan polysulfate sodium; pentostatin; pentrozole;perflubron; perfosfamide; perillyl alcohol; phenazinomycin;phenylacetate; phosphatase inhibitors; picibanil; pilocarpinehydrochloride; pirarubicin; piritrexim; placetin A; placetin B;plasminogen activator inhibitor; platinum complex; platinum compounds;platinum-triamine complex; porfimer sodium; porfiromycin; prednisone;propyl bis-acridone; prostaglandin J2; proteasome inhibitors; proteinA-based immune modulator; protein kinase C inhibitor; protein kinase Cinhibitors, microalgal; protein tyrosine phosphatase inhibitors; purinenucleoside phosphorylase inhibitors; purpurins; pyrazoloacridine;pyridoxylated hemoglobin polyoxyethylene conjugate; raf antagonists;raltitrexed; ramosetron; ras farnesyl protein transferase inhibitors;ras inhibitors; ras-GAP inhibitor; retelliptine demethylated; rhenium Re186 etidronate; rhizoxin; ribozymes; RII retinamide; rohitukine;romurtide; roquinimex; rubiginone B1; ruboxyl; safingol; saintopin;SarCNU; sarcophytol A; sargramostim; Sdi 1 mimetics; semustine;senescence derived inhibitor 1; sense oligonucleotides; signaltransduction inhibitors; sizofuran; sobuzoxane; sodium borocaptate;sodium phenylacetate; solverol; somatomedin binding protein; sonermin;sparfosic acid; spicamycin D; spiromustine; splenopentin; spongistatin1; squalamine; stipiamide; stromelysin inhibitors; sulfinosine;superactive vasoactive intestinal peptide antagonist; suradista;suramin; swainsonine; tallimustine; tamoxifen methiodide; tauromustine;tazarotene; tecogalan sodium; tegafur; tellurapyrylium; telomeraseinhibitors; temoporfin; teniposide; tetrachlorodecaoxide; tetrazomine;thaliblastine; thiocoraline; thrombopoietin; thrombopoietin mimetic;thymalfasin; thymopoietin receptor agonist, thymotrinan; thyroidstimulating hormone; tin ethyl etiopurpurin; tirapazamine; titanocenebichloride; topsentin; toremifene; translation inhibitors; tretinoin;triacetyluridine; triciribine; trimetrexate; triptorelin; tropisetron;turosteride; tyrosine kinase inhibitors; tyrphostins; UBC inhibitors;ubenimex; urogenital sinus-derived growth inhibitory factor; urokinasereceptor antagonists; vapreotide; variolin B; velaresol; veramine;verdins; verteporfin; vinorelbine; vinxaltine; vitaxin; vorozole;zanoterone; zeniplatin; zilascorb; and zinostatin stimalamer.

Specific second active agents include, but are not limited to,2-methoxyestradiol, telomestatin, inducers of apoptosis in multiplemyeloma cells (such as, for example, TRAIL), statins, semaxanib,cyclosporin, etanercept, doxycycline, bortezomib, oblimersen(Genasense®), remicade, docetaxel, celecoxib, melphalan, dexamethasone(Decadron®), steroids, gemcitabine, cisplatinum, temozolomide,etoposide, cyclophosphamide, temodar, carboplatin, procarbazine,gliadel, tamoxifen, topotecan, methotrexate, Arisa®, taxol, taxotere,fluorouracil, leucovorin, irinotecan, xeloda, CPT-11, interferon alpha,pegylated interferon alpha (e.g., PEG INTRON-A), capecitabine,cisplatin, thiotepa, fludarabine, carboplatin, liposomal daunorubicin,cytarabine, doxetaxol, pacilitaxel, vinblastine, IL-2, GM-CSF,dacarbazine, vinorelbine, zoledronic acid, palmitronate, biaxin,busulphan, prednisone, bisphosphonate, arsenic trioxide, vincristine,doxorubicin (Doxil®), paclitaxel, ganciclovir, adriamycin, estramustinesodium phosphate (Emcyt®), sulindac, and etoposide.

4.3 Methods of Treatments and Prevention

Methods of this invention encompass methods of treating, preventingand/or managing various types of cancer, diseases and disordersassociated with, or characterized by, undesired angiogenesis, anddiseases and disorders mediated by PDE 4, which comprise administeringto a patient in need of such treatment, prevention and/or management aneffective amount of a compound of this invention. As used herein, unlessotherwise specified, the term “treating” refers to the administration ofa compound of the invention or other additional active agent after theonset of symptoms of the particular disease or disorder. As used herein,unless otherwise specified, the term “preventing” refers to theadministration prior to the onset of symptoms, particularly to patientsat risk of cancer, and other diseases and disorders associated with, orcharacterized by, undesired angiogenesis. The term “prevention” includesthe inhibition of a symptom of the particular disease or disorder.Patients with familial history of cancer and diseases and disordersassociated with, or characterized by, undesired angiogenesis arepreferred candidates for preventive regimens. As used herein and unlessotherwise indicated, the term “managing” encompasses preventing therecurrence of the particular disease or disorder in a patient who hadsuffered from it, and/or lengthening the time a patient who had sufferedfrom the disease or disorder remains in remission.

As used herein, and unless otherwise specified, the term “effectiveamount” refers to the amount of the compound being administeredsufficient to prevent development of or alleviate to some extent one ormore of the symptoms of the condition or disorder being treated as wellas to alleviate or eradicate the cause of the disease itself.

As used herein, the term “cancer” includes, but is not limited to, solidtumors and blood born tumors. The term “cancer” refers to disease ofskin tissues, organs, blood, and vessels, including, but not limited to,cancers of the bladder, bone or blood, brain, breast, cervix, chest,colon, endrometrium, esophagus, eye, head, kidney, liver, lymph nodes,lung, mouth, neck, ovaries, pancreas, prostate, rectum, stomach, testis,throat, and uterus. The cancer can be primary or metastatic. Specificcancers include, but are not limited to, advanced malignancy,amyloidosis, neuroblastoma, meningioma, atypical meningioma,hemangiopericytoma, multiple brain metastase, glioblastoma multiforms,glioblastoma, brain stem glioma, poor prognosis malignant brain tumor,malignant glioma, recurrent malignant glioma, anaplastic astrocytoma,anaplastic oligodendroglioma, neuroendocrine tumor, rectaladenocarcinoma, Dukes C & D colorectal cancer, unresectable colorectalcarcinoma, metastatic hepatocellular carcinoma, Kaposi's sarcoma,karotype acute myeloblastic leukemia, Hodgkin's lymphoma, non-Hodgkin'slymphoma, cutaneous T-Cell lymphoma, cutaneous B-Cell lymphoma, diffuselarge B-Cell lymphoma, low grade follicular lymphoma, melanoma,including, but not limited to, metastatic melanoma (localized melanoma,including ocular melanoma), malignant mesothelioma, malignant pleuraleffusion mesothelioma syndrome, peritoneal carcinoma, papillary serouscarcinoma, gynecologic sarcoma, soft tissue sarcoma, scelroderma,cutaneous vasculitis, Langerhans cell histiocytosis, leiomyosarcoma,fibrodysplasia ossificans progressive, hormone refractory prostatecancer, resected high-risk soft tissue sarcoma, unrescectablehepatocellular carcinoma, Waldenstrom's macroglobulinemia, myelomaincluding, but not limited to, multiple myeloma, smoldering myeloma andindolent myeloma, fallopian tube cancer, androgen independent prostatecancer, androgen dependent stage IV non-metastatic prostate cancer,hormone-insensitive prostate cancer, chemotherapy-insensitive prostatecancer, carcinoma including, but not limited to, papillary thyroidcarcinoma, follicular thyroid carcinoma, and medullary thyroidcarcinoma, and leiomyoma. In a specific embodiment, the cancer ismetastatic. In another embodiment, the cancer is refractory orresistance to chemotherapy or radiation; in particular, refractory tothalidomide.

As used herein, and unless otherwise specified, the terms “diseases ordisorders associated with, or characterized by, undesired angiogenesis,”“diseases or disorders associated with undesired angiogenesis,” and“diseases or disorders characterized by undesired angiogenesis” refer todiseases, disorders and conditions that are caused, mediated or attendedby undesired, unwanted or uncontrolled angiogenesis, including, but notlimited to, inflammatory diseases, autoimmune diseases, geneticdiseases, allergic diseases, bacterial diseases, ocular neovasculardiseases, choroidal neovascular diseases, and retina neovasculardiseases.

Examples of such diseases or disorders associated with undesiredangiogenesis include, but are not limited to, endometriosis, Crohn'sdisease, heart failure, advanced heart failure, renal impairment,diabetic retinopathy, retinopathy of prematurity, corneal graftrejection, neovascular glaucoma, retrolental fibroplasia, proliferativevitreoretinopathy, trachoma, myopia, optic pits, epidemickeratoconjunctivitis; atopic keratitis, superior limbic keratitis,pterygium keratitis sicca, sjogrens, acne rosacea, phylectenulosis,syphilis, lipid degeneration, bacterial ulcer, fungal ulcer, Herpessimplex infection, Herpes zoster infection, protozoan infection, Kaposisarcoma, Mooren ulcer, Terrien's marginal degeneration, mariginalkeratolysis, myelodysplastic syndrome, rheumatoid arthritis, systemiclupus, polyarteritis, trauma, Wegeners sarcoidosis, scleritis, Steven'sJohnson disease, periphigoid radial keratotomy, sickle cell anemia,sarcoid, pseudoxanthoma elasticum, Pagets disease, vein occlusion,artery occlusion, carotid obstructive disease, chronic uveitis, chronicvitritis, Lyme's disease, Eales disease, Bechet's disease, retinitis,choroiditis, presumed ocular histoplasmosis, Bests disease, Stargartsdisease, pars planitis, chronic retinal detachment, hyperviscositysyndromes, toxoplasmosis, rubeosis, sarcodisis, sclerosis, soriatis,psoriasis, primary sclerosing cholangitis, proctitis, primary biliarysrosis, idiopathic pulmonary fibrosis, alcoholic hepatitis, endotoxemia,toxic shock syndrome, osteoarthritis, retrovirus replication, wasting,meningitis, silica-induced fibrosis, asbestos-induced fibrosis,malignancy-associated hypercalcemia, stroke, circulatory shock,periodontitis, gingivitis, macrocytic anemia, refractory anemia,5q-syndrome, and veterinary disorder caused by feline immunodeficiencyvirus, equine infectious anemia virus, caprine arthritis virus, visnavirus, maedi virus or lenti virus.

In specific embodiments of the invention, diseases or disordersassociated with undesired angiogenesis do not include congestive heartfailure, cardiomyopathy, pulmonary edema, endotoxin-mediated septicshock, acute viral myocarditis, cardiac allograft rejection, myocardialinfarction, HIV, hepatitis, adult respiratory distress syndrome,bone-resorption disease, chronic obstructive pulmonary diseases, chronicpulmonary inflammatory disease, dermatitis, cystic fibrosis, septicshock, sepsis, endotoxic shock, hemodynamic shock, sepsis syndrome, postischemic reperfusion injury, fibrotic disease, cachexia, graftrejection, rheumatoid spondylitis, osteoporosis, ulcerative colitis,inflammatory-bowel disease, multiple sclerosis, systemic lupuserythrematosus, erythema nodosum leprosum in leprosy, radiation damage,asthma, hyperoxic alveolar injury, malaria, mycobacterial infection, andopportunistic infections resulting from HIV.

As used herein, and unless otherwise specified, the term “diseases ordisorders mediated by PDE 4” means a condition or disorder that respondsfavorably to modulation, for example, reduction (e.g., inhibition), ofPDE 4 activity. Favorable responses to PDE 4 modulation includealleviation or abrogation of the disease and/or its attendant symptoms,inhibition of the disease (i.e., arrest or reduction of the developmentof the disease), or its clinical symptoms, and regression of the diseaseor its clinical symptoms. A disease or disorder mediated by PDE 4 may becompletely or partially responsive to PDE 4 modulation. A disease ordisorder mediated by PDE 4 may be associated with inappropriate, e.g.,less than or greater than normal, PDE 4 activity. Inappropriate PDE 4functional activity might arise as the result of PDE 4 expression incells which normally do not express PDE 4, decreased PDE 4 expression(leading to, e.g., lipid and metabolic disorders and diseases) orincreased PDE 4 expression.

As used herein, and unless otherwise specified, the terms “modulate” and“modulation” means that the activity or expression of the molecule(e.g., an enzyme) to be modulated is enhanced or decreased. In someembodiments, the activity or expression of the molecule to be modulatedis enhanced by 10%, 20%, 50%, 100%, or 200% or more, as compared to theactivity or expression of the molecule without the modulation. In otherembodiments, the activity or expression of the molecule to be modulatedis decreased by 10%, 20%, 50%, 70%, 80%, or 90% or more, as compared tothe activity or expression of the molecule without the modulation.

Examples of diseases or disorders mediated by PDE 4 include, but are notlimited to, asthma, atopic dermatitis, depression, reperfusion injury,septic shock, toxic shock, endotoxic shock, adult respiratory distresssyndrome, autoimmune diabetes, diabetes insipidus, multi-infarctdementia, AIDS, cancer, Crohn's disease, multiple sclerosis, cerebralischemia, psoriasis, allograft rejection, restenosis, ulceratiavecolitis, cachexia, cerebral malaria, allergic rhino-conjunctivitis,osteoarthritis, rheumatoid arthritis, chronic obstructive pulmonarydisease (COPD), chronic bronchitis, cosinophilic granuloma, andautoimmune encephalomyelitis.

This invention encompasses methods of treating patients who have beenpreviously treated for cancer, diseases or disorders associated with, orcharacterized by, undesired angiogenesis, or diseases or disordersmediated by PDE 4, but are non-responsive to standard therapies, as wellas those who have not previously been treated. The invention alsoencompasses methods of treating patients regardless of patient's age,although some diseases or disorders are more common in certain agegroups. The invention further encompasses methods of treating patientswho have undergone surgery in an attempt to treat the disease orcondition at issue, as well as those who have not. Because patients withcancer and diseases and disorders characterized by undesiredangiogenesis or mediated by PDE 4 have heterogenous clinicalmanifestations and varying clinical outcomes, the treatment given to apatient may vary, depending on his/her prognosis. The skilled clinicianwill be able to readily determine without undue experimentation specificsecondary agents, types of surgery, and types of non-drug based standardtherapy that can be effectively used to treat an individual patient withcancer and other diseases or disorders.

Methods encompassed by this invention comprise administering one or morecompounds of the invention to a patient (e.g., a human) who issuffering, or likely to suffer, from cancer, a disease or disordermediated by undesired angiogenesis, or a disease or disorder mediated byPDE 4.

In one embodiment of the invention, the recommended daily dose range ofa compound of this invention for the conditions described herein liewithin the range of from about 1 mg to about 10,000 mg per day, given asa single once-a-day dose, or preferably in divided doses throughout aday. More specifically, the daily dose is administered twice daily inequally divided doses. Specifically, a daily dose range should be fromabout 1 mg to about 5,000 mg per day, more specifically, between about10 mg and about 2,500 mg per day, between about 100 mg and about 800 mgper day, between about 100 mg and about 1,200 mg per day, or betweenabout 25 mg and about 2,500 mg per day. In managing the patient, thetherapy should be initiated at a lower dose, perhaps about 1 mg to about2,500 mg, and increased if necessary up to about 200 mg to about 5,000mg per day as either a single dose or divided doses, depending on thepatient's global response.

4.3.1 Combination Therapy with a Second Active Agent

Specific methods of the invention comprise administering a compound ofthis invention, or a pharmaceutically acceptable salt, solvate, hydrate,stereoisomer, clathrate, or prodrug thereof, in combination with one ormore second active agents, and/or in combination with radiation therapy,blood transfusions, or surgery. Examples of second active agents arealso disclosed herein (see, e.g., section 4.2).

Administration of the compound of this invention and the second activeagents to a patient can occur simultaneously or sequentially by the sameor different routes of administration. The suitability of a particularroute of administration employed for a particular active agent willdepend on the active agent itself (e.g., whether it can be administeredorally without decomposing prior to entering the blood stream) and thedisease being treated. A preferred route of administration for acompound of this invention of the invention is oral or ophthalmic.Preferred routes of administration for the second active agents oringredients of the invention are known to those of ordinary skill in theart. See, e.g., Physicians'Desk Reference, 1755-1760 (56^(th) ed.,2002).

In one embodiment of the invention, the second active agent isadministered intravenously or subcutaneously and once or twice daily inan amount of from about 1 to about 1000 mg, from about 5 to about 500mg, from about 10 to about 350 mg, or from about 50 to about 200 mg. Thespecific amount of the second active agent will depend on the specificagent used, the type of disease being treated or managed, the severityand stage of disease, and the amount(s) of compounds of the inventionand any optional additional active agents concurrently administered tothe patient. In a particular embodiment, the second active agent isoblimersen (Genasense®), GM-CSF, G-CSF, EPO, taxotere, irinotecan,dacarbazine, transretinoic acid, topotecan, pentoxifylline,ciprofloxacin, dexamethasone, vincristine, doxorubicin, cox-2inhibitors, IL2, IL8, IL18, IFN, Ara-C, vinorelbine, or a combinationthereof.

In a particular embodiment, GM-CSF, G-CSF or EPO is administeredsubcutaneously during about five days in a four or six week cycle in anamount of from about 1 to about 750 mg/m²/day, preferably in an amountof from about 25 to about 500 mg/m²/day, more preferably in an amount offrom about 50 to about 250 mg/m²/day, and most preferably in an amountof from about 50 to about 200 mg/m²/day. In a certain embodiment, GM-CSFmay be administered in an amount of from about 60 to about 500 mcg/m²intravenously over 2 hours, or from about 5 to about 12 mcg/m²/daysubcutaneously. In a specific embodiment, G-CSF may be administeredsubcutaneously in an amount of about 1 mcg/kg/day initially and can beadjusted depending on rise of total granulocyte counts. The maintenancedose of G-CSF may be administered in an amount of about 300 (in smallerpatients) or 480 mcg subcutaneously. In a certain embodiment, EPO may beadministered subcutaneously in an amount of 10,000 Unit 3 times perweek.

In another embodiment, a compound of this invention is administered inan amount of from about 20 mg to about 1,200 mg/d alone or incombination with a second active agent to patients with metastaticmelanoma (localized melanoma, including, but not limited to, ocularmelanoma). In one embodiment, a compound of this invention in an amountof from about 800 to about 1,200 mg/d and dacarbazine (DTIC) in anamount of from about 200 to about 1000 mg/m²/d are administered topatients with metastatic melanoma (localized melanoma, including, butnot limited to, ocular melanoma). In another embodiment, a compound ofthis invention in an amount of about from 800 to 1,200 mg/d andtemozolomide are administered to patients with metastatic melanoma(localized melanoma, including, but not limited to, ocular-melanoma). Inanother embodiment, a compound of this invention is administered topatients with metastatic melanoma or localized melanoma whose diseasehas progressed on treatment with temozolomide, dacarbazine (DTIC), IL-2and/or IFN. In a specific embodiment, a compound of this invention isadministered to patients with relapsed or refractory multiple myeloma incombination with dexamethasone.

In another embodiment, a compound of this invention is administered withmelphalan and dexamethasone to patients with amyloidosis. In a specificembodiment, a compound of the invention and steroids can be administeredto patients with amyloidosis.

In another embodiment, a compound of this invention is administered withgemcitabine and cisplatinum to patients with locally advanced ormetastatic transitional cell bladder cancer.

In another embodiment, a compound of this invention is administered incombination with a second active ingredient as follows: temozolomide topediatric patients with relapsed or progressive brain tumors orrecurrent neuroblastoma; celecoxib, etoposide and cyclophosphamide forrelapsed or progressive CNS cancer; temodar to patients with recurrentor progressive meningioma, malignant meningioma, hemangiopericytoma,multiple brain metastases, relapased brain tumors, or newly diagnosedglioblastoma multiforms; irinotecan to patients with recurrentglioblastoma; carboplatin to pediatric patients with brain stem glioma;procarbazine to pediatric patients with progressive malignant gliomas;cyclophosphamide to patients with poor prognosis malignant brain tumors,newly diagnosed or recurrent glioblastoma multiforms; Gliadel® for highgrade recurrent malignant gliomas; temozolomide and tamoxifen foranaplastic astrocytoma; or topotecan for gliomas, glioblastoma,anaplastic astrocytoma or anaplastic oligodendroglioma.

In another embodiment, a compound of this invention is administered withmethotrexate and cyclophosphamide to patients with metastatic breastcancer.

In another embodiment, a compound of this invention is administered withtemozolomide to patients with neuroendocrine tumors.

In another embodiment, a compound of this invention is administered withgemcitabine to patients with recurrent or metastatic head or neckcancer. In another embodiment, a compound of this invention isadministered with gemcitabine to patients with pancreatic cancer.

In another embodiment, a compound of this invention is administered topatients with colon cancer in combination with Arisa®, taxol and/ortaxotere.

In another embodiment, a compound of this invention is administered withcapecitabine to patients with refractory colorectal cancer or patientswho fail first line therapy or have poor performance in colon or rectaladenocarcinoma.

In another embodiment, a compound of this invention is administered incombination with fluorouracil, leucovorin, and irinotecan to patientswith Dukes C & D colorectal cancer or to patients who have beenpreviously treated for metastatic colorectal cancer.

In another embodiment, a compound of this invention is administered topatients with refractory colorectal cancer in combination withcapecitabine, xeloda, and/or CPT-11.

In another embodiment, a compound of the invention is administered withcapecitabine and irinotecan to patients with refractory colorectalcancer or to patients with unresectable or metastatic colorectalcarcinoma.

In another embodiment, a compound of this invention is administeredalone or in combination with interferon alpha or capecitabine topatients with unresectable or metastatic hepatocellular carcinoma; orwith cisplatin and thiotepa to patients with primary or metastatic livercancer.

In another embodiment, a compound of this invention is administered incombination with pegylated interferon alpha to patients with Kaposi'ssarcoma.

In another embodiment, a compound of this invention is administered incombination with fludarabine, carboplatin, and/or topotecan to patientswith refractory or relapsed or high-risk acuted myelogenous leukemia.

In another embodiment, a compound of this invention is administered incombination with liposomal daunorubicin, topotecan and/or cytarabine topatients with unfavorable karotype acute myeloblastic leukemia.

In another embodiment, a compound of this invention is administered incombination with gemcitabine and irinotecan to patients with non-smallcell lung cancer. In one embodiment, a compound of this invention isadministered in combination with carboplatin and irinotecan to patientswith non-small cell lung cancer. In one embodiment, a compound of thisinvention is administered with doxetaxol to patients with non-small celllung cancer who have been previously treated with carbo/VP 16 andradiotherapy.

In another embodiment, a compound of this invention is administered incombination with carboplatin and/or taxotere, or in combination withcarboplatin, pacilitaxel and/or thoracic radiotherapy to patients withnon-small cell lung cancer. In a specific embodiment, a compound of thisinvention is administered in combination with taxotere to patients withstage IIIB or IV non-small cell lung cancer.

In another embodiment, a compound of this invention is administered incombination with oblimersen (Genasense®) to patients with small celllung cancer.

In another embodiment, a compound of this invention is administeredalone or in combination with a second active ingredient such asvinblastine or fludarabine to patients with various types of lymphoma,including, but not limited to, Hodgkin's lymphoma, non-Hodgkin'slymphoma, cutaneous T-Cell lymphoma, cutaneous B-Cell lymphoma, diffuselarge B-Cell lymphoma or relapsed or refractory low grade follicularlymphoma.

In another embodiment, a compound of this invention is administered incombination with taxotere, IL-2, IFN, GM-CSF, and/or dacarbazine topatients with various types or stages of melanoma including, but notlimited to, localized melanoma or metastatic melanoma such as ocularmelanoma.

In another embodiment, a compound of this invention is administeredalone or in combination with vinorelbine to patients with malignantmesothelioma, or stage IIIB non-small cell lung cancer with pleuralimplants or malignant pleural effusion mesothelioma syndrome.

In another embodiment, a compound of this invention is administered topatients with various types or stages of multiple myeloma in combinationwith dexamethasone, zoledronic acid, palmitronate, GM-CSF, biaxin,vinblastine, melphalan, busulphan, cyclophosphamide, IFN, palmidronate,prednisone, bisphosphonate, celecoxib, arsenic trioxide, PEG INTRON-A,vincristine, doxil, decadron, or a combination thereof.

In another embodiment, a compound of this invention is administered topatients with relapsed or refractory multiple myeloma in combinationwith doxorubicin (Doxil®), vincristine and/or dexamethasone (Decadron®).

In another embodiment, a compound of this invention is administered topatients with various types or stages of ovarian cancer such asperitoneal carcinoma, papillary serous carcinoma, refractory ovariancancer or recurrent ovarian cancer, in combination with taxol,carboplatin, doxorubicin, gemcitabine, cisplatin, xeloda, paclitaxel,dexamethasone, or a combination thereof.

In another embodiment, a compound of this invention is administered topatients with various types or stages of prostate cancer, in combinationwith xeloda, 5 FU/LV, gemcitabine, irinotecan plus gemcitabine,cyclophosphamide, vincristine, dexamethasone, GM-CSF, celecoxib,taxotere, ganciclovir, paclitaxel, adriamycin, docetaxel, estramustine,Emcyt, or a combination thereof.

In another embodiment, a compound of this invention is administered topatients with various types or stages of renal cell cancer, incombination with capecitabine, IFN, tamoxifen, IL-2, GM-CSF, Celebrex®,or a combination thereof.

In another embodiment, a compound of this invention is administered topatients with various types or stages of gynecologic, uterus or softtissue sarcoma cancer in combination with IFN, a COX-2 inhibitor such asCelebrex®, and/or sulindac.

In another embodiment, a compound of this invention is administered topatients with various types or stages of solid tumors in combinationwith celebrex, etoposide, cyclophosphamide, docetaxel, apecitabine, IFN,tamoxifen, IL-2, GM-CSF, or a combination thereof.

In another embodiment, a compound of this invention is administered topatients with scelroderma or cutaneous vasculitis in combination withcelebrex, etoposide, cyclophosphamide, docetaxel, apecitabine, IFN,tamoxifen, IL-2, GM-CSF, or a combination thereof.

This invention also encompasses a method of increasing the dosage of ananti-cancer drug or agent that can be safely and effectivelyadministered to a patient, which comprises administering to a patient(e.g., a human) a compound of this invention. Patients that can benefitby this method are those likely to suffer from an adverse effectassociated with anti-cancer drugs for treating a specific cancer of theskin, subcutaneous tissue, lymph nodes, brain, lung, liver, bone,intestine, colon, heart, pancreas, adrenal, kidney, prostate, breast,colorectal, or combinations thereof. The administration of a compound ofthis invention alleviates or reduces adverse effects which are of suchseverity that it would otherwise limit the amount of anti-cancer drug.

In one embodiment, a compound of this invention can be administeredorally and daily in an amount of from about 1 to about 5,000 mg, fromabout 10 to about 2,500 mg, from about 25 to about 2,500 mg, from about100 to about 1,200 mg, or from about 100 to about 800 mg prior to,during, or after the occurrence of the adverse effect associated withthe administration of an anti-cancer drug to a patient. In a particularembodiment, a compound of this invention is administered in combinationwith specific agents such as heparin, aspirin, coumadin, or G-CSF toavoid adverse effects that are associated with anti-cancer drugs such asbut not limited to neutropenia or thrombocytopenia.

In one embodiment, a compound of this invention can be administered topatients with diseases and disorders associated with, or characterizedby, undesired angiogenesis, optionally in combination with additionalactive ingredients including but not limited to anti-cancer drugs,anti-inflammatories, antihistamines, antibiotics, and steroids.

In another embodiment, this invention encompasses a method of treating,preventing and/or managing cancer, which comprises administering acompound of this invention, optionally in conjunction with (e.g. before,during, or after) conventional therapy including, but not limited to,surgery, immunotherapy, biological therapy, radiation therapy, or othernon-drug based therapy presently used to treat, prevent or managecancer. The combined use of the compounds of this invention andconventional therapy may provide a unique treatment regimen that isunexpectedly effective in certain patients. Without being limited bytheory, it is believed that compound of this invention may provideadditive or synergistic effects when given concurrently withconventional therapy.

In another embodiment, this invention encompasses a method of treating,preventing and/or managing diseases and disorders associated with, orcharacterized by, undesired angiogenesis, which comprises administeringa compound of this invention, optionally in conjunction with (e.g.before, during, or after) conventional therapy including, but notlimited to, surgery, immunotherapy, biological therapy, radiationtherapy, or other non-drug based therapy presently used to treat,prevent or manage diseases and disorders associated with, orcharacterized by, undesired angiogenesis. The combined use of thecompounds of this invention and conventional therapy may provide aunique treatment regimen that is unexpectedly effective in certainpatients. Without being limited by theory, it is believed that thecompounds of this invention may provide additive or synergistic effectswhen given concurrently with conventional therapy.

As discussed elsewhere herein, the invention encompasses a method ofreducing, treating and/or preventing adverse or undesired effectsassociated with conventional therapy including, but not limited to,surgery, chemotherapy, radiation therapy, hormonal therapy, biologicaltherapy and immunotherapy. One or more compounds of this invention,optionally with other active ingredient, can be administered to apatient prior to, during, or after the occurrence of the adverse effectassociated with conventional therapy.

In one embodiment, a compound of this invention can be administered inan amount of from about 1 to about 5,000 mg, from about 10 to about2,500 mg, from about 25 to about 2,500 mg, from about 100 to about 1,200mg, or from about 100 to about 800 mg orally and daily alone, or incombination with a second active agent disclosed herein (see, e.g.,section 4.2), prior to, during, or after the use of conventionaltherapy.

In a specific embodiment of this method, a compound of this inventionand doxetaxol are administered to patients with non-small cell lungcancer who were previously treated with carbo/VP 16 and radiotherapy.

4.3.2 Use with Transplantation Therapy

Compounds of the invention can be used to reduce the risk of GraftVersus Host Disease (GVHD). Therefore, the invention encompasses amethod of treating, preventing and/or managing cancer, which comprisesadministering a compound of this invention, optionally in conjunctionwith transplantation therapy.

As those of ordinary skill in the art are aware, the treatment of canceris often based on the stages and mechanism of the disease. For example,as inevitable leukemic transformation develops in certain stages ofcancer, transplantation of peripheral blood stem cells, hematopoieticstem cell preparation or bone marrow may be necessary. The combined useof the compound of this invention and transplantation therapy provides aunique and unexpected synergism. In particular, a compound of thisinvention exhibits activity that may provide additive or synergisticeffects when given concurrently with transplantation therapy in patientswith cancer.

A compound of this invention can work in combination withtransplantation therapy reducing complications associated with theinvasive procedure of transplantation and risk of GVHD. This inventionencompasses a method of treating, preventing and/or managing cancerwhich comprises administering to a patient (e.g., a human) a compound ofthis invention, before, during, or after the transplantation ofumbilical cord blood, placental blood, peripheral blood stem cell,hematopoietic stem cell preparation or bone marrow. Examples of stemcells suitable for use in the methods of the invention are disclosed inU.S. patent application Ser. No. 10/411,655, filed Apr. 11, 2003 by R.Hariri et al., the entirety of which is incorporated herein byreference.

In another embodiment, this invention encompasses a method of treating,preventing and/or managing diseases and disorders associated with, orcharacterized by, undesired angiogenesis, which comprises administeringto a patient (e.g., a human) a compound of this invention, before,during, or after the transplantation of umbilical cord blood, placentalblood, peripheral blood stem cell, hematopoietic stem cell preparationor bone marrow.

In one embodiment of this method, a compound of this invention isadministered to patients with multiple myeloma before, during, or afterthe transplantation of autologous peripheral blood progenitor cell.

In another embodiment, a compound of this invention is administered topatients with relapsing multiple myeloma after the stem celltransplantation.

In another embodiment, a compound of this invention and prednisone areadministered as maintenance therapy to patients with multiple myelomafollowing the transplantation of autologous stem cell.

In another embodiment, a compound of this invention and dexamethasoneare administered as salvage therapy for low risk post transplantation topatients with multiple myeloma.

In another embodiment, a compound of this invention and dexamethasoneare administered as maintenance therapy to patients with multiplemyeloma following the transplantation of autologous bone marrow.

In another embodiment, a compound of this invention is administeredfollowing the administration of high dose of melphalan and thetransplantation of autologous stem cell to patients with chemotherapyresponsive multiple myeloma.

In another embodiment, a compound of this invention and PEG INTRO-A areadministered as maintenance therapy to patients with multiple myelomafollowing the transplantation of autologous CD34-selected peripheralstem cell.

In another embodiment, a compound of this invention is administered withpost transplant consolidation chemotherapy to patients with newlydiagnosed multiple myeloma to evaluate anti-angiogenesis.

In another embodiment, a compound of this invention and dexamethasoneare administered as maintenance therapy after DCEP consolidation,following the treatment with high dose of melphalan and thetransplantation of peripheral blood stem cell to 65 years of age orolder patients with multiple myeloma.

4.3.3 Cycling Therapy

In certain embodiments, the prophylactic or therapeutic agents of theinvention are cyclically administered to a patient. Cycling therapyinvolves the administration of an active agent for a period of time,followed by a rest for a period of time, and repeating this sequentialadministration. Cycling therapy can reduce the development of resistanceto one or more of the therapies, avoid or reduce the side effects of oneof the therapies, and/or improves the efficacy of the treatment.

Consequently, in one specific embodiment of the invention, a compound ofthis invention is administered daily in a single or divided doses in afour to six week cycle with a rest period of about a week or two weeks.The invention further allows the frequency, number, and length of dosingcycles to be increased. Thus, another specific embodiment of theinvention encompasses the administration of a compound of this inventionfor more cycles than are typical when it is administered alone. In yetanother specific embodiment of the invention, a compound of thisinvention is administered for a greater number of cycles that wouldtypically cause dose-limiting toxicity in a patient to whom a secondactive ingredient is not also being administered.

In one embodiment, a compound of this invention is administered dailyand continuously for three or four weeks at a dose of from about 1 toabout 5,000 mg/d followed by a break of one or two weeks. A compound ofthis invention is preferably administered daily and continuously at aninitial dose of 1 to 5 mg/d with dose escalation (every week) by 10 to100 mg/d to a maximum dose of 5,000 mg/d for as long as therapy istolerated. In a particular embodiment, the compound is administered inan amount of about 400, 800, or 1,200 mg/day, preferably in an amount ofabout 800 mg/day for three to four weeks, followed by one week or twoweeks of rest in a four or six week cycle.

In one embodiment of the invention, a compound of this invention and asecond active ingredient are administered orally, with administration ofa compound of this invention occurring 30 to 60 minutes prior to asecond active ingredient, during a cycle of four to six weeks. Inanother embodiment of the invention, the combination of a compound ofthis invention and a second active ingredient is administered byintravenous infusion over about 90 minutes every cycle. In a specificembodiment, one cycle comprises the administration of from about 400 toabout 800 mg/day of a compound of this invention and from about 50 toabout 200 mg/m²/day of a second active ingredient daily for 3 to 4 weeksand then one or two weeks of rest. In another specific embodiment, eachcycle comprises the administration of from about 200 to about 400 mg/dayof a compound of this invention and from about 50 to about 200 mg/m²/dayof a second active ingredient for three to four weeks followed by one ortwo weeks of rest. Typically, the number of cycles during which thecombinatorial treatment is administered to a patient will be from aboutone to about 24 cycles, more typically from about two to about 16cycles, and even more typically from about four to about eight cycles.

4.4 Pharmaceutical Compositions

Pharmaceutical compositions can be used in the preparation ofindividual, single unit dosage forms. Pharmaceutical compositions anddosage forms of the invention comprise a compound of this invention.Pharmaceutical compositions and dosage forms of the invention canfurther comprise one or more excipients.

Pharmaceutical compositions and dosage forms of the invention can alsocomprise one or more additional active ingredients. Consequently,pharmaceutical compositions and dosage forms of the invention comprisethe active ingredients disclosed herein (e.g., a compound of thisinvention and optionally a second active agent). Examples of optionalsecond, or additional, active ingredients are disclosed herein (see,e.g., section 4.2).

Single unit dosage forms of the invention are suitable for oral, mucosal(e.g. nasal, sublingual, vaginal, buccal, or rectal), parenteral (e.g.,subcutaneous, intravenous, bolus injection, intramuscular, orintraarterial), topical (e.g., eye drops or other ophthalmicpreparations), transdermal or transcutaneous administration to apatient. Examples of dosage forms include, but are not limited to:tablets; caplets; capsules, such as soft elastic gelatin capsules;cachets; troches; lozenges; dispersions; suppositories; powders;aerosols (e.g., nasal sprays or inhalers); gels; liquid dosage formssuitable for oral or mucosal administration to a patient, includingsuspensions (e.g., aqueous or non-aqueous liquid suspensions,oil-in-water emulsions, or a water-in-oil liquid emulsions), solutions,and elixirs; liquid dosage forms suitable for parenteral administrationto a patient; eye drops or other ophthalmic preparations suitable fortopical administration; and sterile solids (e.g., crystalline oramorphous solids) that can be reconstituted to provide liquid dosageforms suitable for parenteral administration to a patient.

The composition, shape, and type of dosage forms of the invention willtypically vary depending on their use. For example, a dosage form usedin the acute treatment of a disease may contain larger amounts of one ormore of the active ingredients it comprises than a dosage form used inthe chronic treatment of the same disease. Similarly, a parenteraldosage form may contain smaller amounts of one or more of the activeingredients it comprises than an oral dosage form used to treat the samedisease. These and other ways in which specific dosage forms encompassedby this invention will vary from one another will be readily apparent tothose skilled in the art. See, e.g., Remington's PharmaceuticalSciences, 18th ed., Mack Publishing, Easton Pa. (1990).

Typical pharmaceutical compositions and dosage forms comprise one ormore excipients. Suitable excipients are well known to those skilled inthe art of pharmacy, and non-limiting examples of suitable excipientsare provided herein. Whether a particular excipient is suitable forincorporation into a pharmaceutical composition or dosage form dependson a variety of factors well known in the art including, but not limitedto, the way in which the dosage form will be administered to a patient.For example, oral dosage forms such as tablets may contain excipientsnot suited for use in parenteral dosage forms. The suitability of aparticular excipient may also depend on the specific active ingredientsin the dosage form. For example, the decomposition of some activeingredients may be accelerated by some excipients such as lactose, orwhen exposed to water. Active ingredients that comprise primary orsecondary amines are particularly susceptible to such accelerateddecomposition. Consequently, this invention encompasses pharmaceuticalcompositions and dosage forms that contain little, if any, lactose othermono- or di-saccharides. As used herein, the term “lactose-free” meansthat the amount of lactose present, if any, is insufficient tosubstantially increase the degradation rate of an active ingredient.

Lactose-free compositions of the invention can comprise excipients thatare well known in the art and are listed, for example, in the U.S.Pharmacopeia (USP) 25-NF20 (2002). In general, lactose-free compositionscomprise active ingredients, a binder/filler, and a lubricant inpharmaceutically compatible and pharmaceutically acceptable amounts.Preferred lactose-free dosage forms comprise active ingredients,microcrystalline cellulose, pre-gelatinized starch, and magnesiumstearate.

This invention further encompasses anhydrous pharmaceutical compositionsand dosage forms comprising active ingredients, since water canfacilitate the degradation of some compounds. For example, the additionof water (e.g., 5%) is widely accepted in the pharmaceutical arts as ameans of simulating long-term storage in order to determinecharacteristics such as shelf-life or the stability of formulations overtime. See, e.g., Jens T. Carstensen, Drug Stability: Principles &Practice, 2d. Ed., Marcel Dekker, NY, N.Y., 1995, pp. 379-80. In effect,water and heat accelerate the decomposition of some compounds. Thus, theeffect of water on a formulation can be of great significance sincemoisture and/or humidity are commonly encountered during manufacture,handling, packaging, storage, shipment, and use of formulations.

Anhydrous pharmaceutical compositions and dosage forms of the inventioncan be prepared using anhydrous or low moisture containing ingredientsand low moisture or low humidity conditions. Pharmaceutical compositionsand dosage forms that comprise lactose and at least one activeingredient that comprises a primary or secondary amine are preferablyanhydrous if substantial contact with moisture and/or humidity duringmanufacturing, packaging, and/or storage is expected.

An anhydrous pharmaceutical composition should be prepared and storedsuch that its anhydrous nature is maintained. Accordingly, anhydrouscompositions are preferably packaged using materials known to preventexposure to water such that they can be included in suitable formularykits. Examples of suitable packaging include, but are not limited to,hermetically sealed foils, plastics, unit dose containers (e.g., vials),blister packs, and strip packs.

The invention further encompasses pharmaceutical compositions and dosageforms that comprise one or more compounds that reduce the rate by whichan active ingredient will decompose. Such compounds, which are referredto herein as “stabilizers,” include, but are not limited to,antioxidants such as ascorbic acid, pH buffers, or salt buffers.

Like the amounts and types of excipients, the amounts and specific typesof active ingredients in a dosage form may differ depending on factorssuch as, but not limited to, the route by which it is to be administeredto patients. However, typical dosage forms of the invention comprise acompound of this invention in an amount of from about 0.10 to about 150mg. Typical dosage forms comprise a compound of this invention in anamount of about 0.1, 1, 2, 5, 7.5, 10, 12.5, 15, 17.5, 20, 25, 50, 100,150 or 200 mg. In a specific embodiment, a preferred dosage formcomprises a compound of this invention in an amount of about 5, 10, 25or 50 mg. Typical dosage forms comprise the second active ingredient inan amount of 1 to about 1000 mg, from about 5 to about 500 mg, fromabout 10 to about 350 mg, or from about 50 to about 200 mg. Of course,the specific amount of the anti-cancer drug will depend on the specificagent used, the type of cancer being treated or managed, and theamount(s) of a compound of this invention and any optional additionalactive agents concurrently administered to the patient.

4.4.1 Oral Dosage Forms

Pharmaceutical compositions of the invention that are suitable for oraladministration can be presented as discrete dosage forms, such as, butare not limited to, tablets (e.g., chewable tablets), caplets, capsules,and liquids (e.g., flavored syrups). Such dosage forms containpredetermined amounts of active ingredients, and may be prepared bymethods of pharmacy well known to those skilled in the art. Seegenerally, Remington's Pharmaceutical Sciences, 18th ed., MackPublishing, Easton Pa. (1990).

Typical oral dosage forms of the invention are prepared by combining theactive ingredients in an intimate admixture with at least one excipientaccording to conventional pharmaceutical compounding techniques.Excipients can take a wide variety of forms depending on the form ofpreparation desired for administration. For example, excipients suitablefor use in oral liquid or aerosol dosage forms include, but are notlimited to, water, glycols, oils, alcohols, flavoring agents,preservatives, and coloring agents. Examples of excipients suitable foruse in solid oral dosage forms (e.g., powders, tablets, capsules, andcaplets) include, but are not limited to, starches, sugars,micro-crystalline cellulose, diluents, granulating agents, lubricants,binders, and disintegrating agents.

Because of their-ease of administration, tablets and capsules representthe most advantageous oral dosage unit forms, in which case solidexcipients are employed. If desired, tablets can be coated by standardaqueous or nonaqueous techniques. Such dosage forms can be prepared byany of the methods of pharmacy. In general, pharmaceutical compositionsand dosage forms are prepared by uniformly and intimately admixing theactive ingredients with liquid carriers, finely divided solid carriers,or both, and then shaping the product into the desired presentation ifnecessary.

For example, a tablet can be prepared by compression or molding.Compressed tablets can be prepared by compressing in a suitable machinethe active ingredients in a free-flowing form such as powder orgranules, optionally mixed with an excipient. Molded tablets can be madeby molding in a suitable machine a mixture of the powdered compoundmoistened with an inert liquid diluent.

Examples of excipients that can be used in oral dosage forms of theinvention include, but are not limited to, binders, fillers,disintegrants, and lubricants. Binders suitable for use inpharmaceutical compositions and dosage forms include, but are notlimited to, corn starch, potato starch, or other starches, gelatin,natural and synthetic gums such as acacia, sodium alginate, alginicacid, other alginates, powdered tragacanth, guar gum, cellulose and itsderivatives (e.g., ethyl cellulose, cellulose acetate, carboxymethylcellulose calcium, sodium carboxymethyl cellulose), polyvinylpyrrolidone, methyl cellulose, pre-gelatinized starch, hydroxypropylmethyl cellulose, (e.g., Nos. 2208, 2906, 2910), microcrystallinecellulose, and mixtures thereof.

Suitable forms of microcrystalline cellulose include, but are notlimited to, the materials sold as AVICEL-PH-101, AVICEL-PH-103 AVICELRC-581, AVICEL-PH-105 (available from FMC Corporation, American ViscoseDivision, Avicel Sales, Marcus Hook, Pa.), and mixtures thereof. Anspecific binder is a mixture of microcrystalline cellulose and sodiumcarboxymethyl cellulose sold as AVICEL RC-581. Suitable anhydrous or lowmoisture excipients or additives include AVICEL-PH-103™ and Starch 1500LM.

Examples of fillers suitable for use in the pharmaceutical compositionsand dosage forms disclosed herein include, but are not limited to, talc,calcium carbonate (e.g., granules or powder), microcrystallinecellulose, powdered cellulose, dextrates, kaolin, mannitol, silicicacid, sorbitol, starch, pre-gelatinized starch, and mixtures thereof.The binder or filler in pharmaceutical compositions of the invention istypically present in from about 50 to about 99 weight percent of thepharmaceutical composition or dosage form.

Disintegrants are used in the compositions of the invention to providetablets that disintegrate when exposed to an aqueous environment.Tablets that contain too much disintegrant may disintegrate in storage,while those that contain too little may not disintegrate at a desiredrate or under the desired conditions. Thus, a sufficient amount ofdisintegrant that is neither too much nor too little to detrimentallyalter the release of the active ingredients should be used to form solidoral dosage forms of the invention. The amount of disintegrant usedvaries based upon the type of formulation, and is readily discernible tothose of ordinary skill in the art. Typical pharmaceutical compositionscomprise from about 0.5 to about 15 weight percent of disintegrant,preferably from about 1 to about 5 weight percent of disintegrant.

Disintegrants that can be used in pharmaceutical compositions and dosageforms of the invention include, but are not limited to, agar-agar,alginic acid, calcium carbonate, microcrystalline cellulose,croscarmellose sodium, crospovidone, polacrilin potassium, sodium starchglycolate, potato or tapioca starch, other starches, pre-gelatinizedstarch, other starches, clays, other algins, other celluloses, gums, andmixtures thereof.

Lubricants that can be used in pharmaceutical compositions and dosageforms of the invention include, but are not limited to, calciumstearate, magnesium stearate, mineral oil, light mineral oil, glycerin,sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid,sodium lauryl sulfate, talc, hydrogenated vegetable oil (e.g., peanutoil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, andsoybean oil), zinc stearate, ethyl oleate, ethyl laureate, agar, andmixtures thereof. Additional lubricants include, for example, a syloidsilica gel (AEROSIL200, manufactured by W. R. Grace Co. of Baltimore,Md.), a coagulated aerosol of synthetic silica (marketed by Degussa Co.of Piano, Tex.), CAB-O-SIL (a pyrogenic silicon dioxide product sold byCabot Co. of Boston, Mass.), and mixtures thereof. If used at all,lubricants are typically used in an amount of less than about 1 weightpercent of the pharmaceutical compositions or dosage forms into whichthey are incorporated.

A preferred solid oral dosage form of the invention comprises a compoundof this invention, anhydrous lactose, microcrystalline cellulose,polyvinylpyrrolidone, stearic acid, colloidal anhydrous silica, andgelatin.

4.4.2 Delayed Release Dosage Forms

Active ingredients of the invention can be administered by controlledrelease means or by delivery devices that are well known to those ofordinary skill in the art. Examples include, but are not limited to,those described in U.S. Pat. Nos. 3,845,770; 3,916,899; 3,536,809;3,598,123; and 4,008,719, 5,674,533, 5,059,595, 5,591,767, 5,120,548,5,073,543, 5,639,476, 5,354,556, and 5,733,566, each of which isincorporated herein by reference. Such dosage forms can be used toprovide slow or controlled-release of one or more active ingredientsusing, for example, hydropropylmethyl cellulose, other polymer matrices,gels, permeable membranes, osmotic systems, multilayer coatings,microparticles, liposomes, microspheres, or a combination thereof toprovide the desired release profile in varying proportions. Suitablecontrolled-release formulations known to those of ordinary skill in theart, including those described herein, can be readily selected for usewith the active ingredients of the invention. The invention thusencompasses single unit dosage forms suitable for oral administrationsuch as, but not limited to, tablets, capsules, gelcaps, and capletsthat are adapted for controlled-release.

All controlled-release pharmaceutical products have a common goal ofimproving drug therapy over that achieved by their non-controlledcounterparts. Ideally, the use of an optimally designedcontrolled-release preparation in medical treatment is characterized bya minimum of drug substance being employed to cure or control thecondition in a minimum amount of time. Advantages of controlled-releaseformulations include extended activity of the drug, reduced dosagefrequency, and increased patient compliance. In addition,controlled-release formulations can be used to affect the time of onsetof action or other characteristics, such as blood levels of the drug,and can thus affect the occurrence of side (e.g., adverse) effects.

Most controlled-release formulations are designed to initially releasean amount of drug (active ingredient) that promptly produces the desiredtherapeutic effect, and gradually and continually release of otheramounts of drug to maintain this level of therapeutic or prophylacticeffect over an extended period of time. In order to maintain thisconstant level of drug in the body, the drug must be released from thedosage form at a rate that will replace the amount of drug beingmetabolized and excreted from the body. Controlled-release of an activeingredient can be stimulated by various conditions including, but notlimited to, pH, temperature, enzymes, water, or other physiologicalconditions or compounds.

4.4.3 Parenteral Dosage Forms

Parenteral dosage forms can be administered to patients by variousroutes including, but not limited to, subcutaneous, intravenous(including bolus injection), intramuscular, and intraarterial. Becausetheir administration typically bypasses patients' natural defensesagainst contaminants, parenteral dosage forms are preferably sterile orcapable of being sterilized prior to administration to a patient.Examples of parenteral dosage forms include, but are not limited to,solutions ready for injection, dry products ready to be dissolved orsuspended in a pharmaceutically acceptable vehicle for injection,suspensions ready for injection, and emulsions.

Suitable vehicles that can be used to provide parenteral dosage forms ofthe invention are well known to those skilled in the art. Examplesinclude, but are not limited to: Water for Injection USP; aqueousvehicles such as, but not limited to, Sodium Chloride Injection,Ringer's Injection, Dextrose Injection, Dextrose and Sodium ChlorideInjection, and Lactated Ringer's Injection; water-miscible vehicles suchas, but not limited to, ethyl alcohol, polyethylene glycol, andpolypropylene glycol; and non-aqueous vehicles such as, but not limitedto, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate,isopropyl myristate, and benzyl benzoate.

Compounds that increase the solubility of one or more of the activeingredients disclosed herein can also be incorporated into theparenteral dosage forms of the invention. For example, cyclodextrin andits derivatives can be used to increase the solubility of a compound ofthis invention. See, e.g., U.S. Pat. No. 5,134,127, which isincorporated herein by reference.

4.4.4 Topical and Mucosal Dosage Forms

Topical and mucosal dosage forms of the invention include, but are notlimited to, sprays, aerosols, solutions, emulsions, suspensions, eyedrops or other ophthalmic preparations, or other forms known to one ofskill in the art. See, e.g., Remington's Pharmaceutical Sciences,16^(th) and 18^(th) eds., Mack Publishing, Easton Pa. (1980 & 1990); andIntroduction to Pharmaceutical Dosage Forms, 4th ed., Lea & Febiger,Philadelphia (1985). Dosage forms suitable for treating mucosal tissueswithin the oral cavity can be formulated as mouthwashes or as oral gels.

Suitable excipients (e.g., carriers and diluents) and other materialsthat can be used to provide topical and mucosal dosage forms encompassedby this invention are well known to those skilled in the pharmaceuticalarts, and depend on the particular tissue to which a givenpharmaceutical composition or dosage form will be applied. With thatfact in mind, typical excipients include, but are not limited to, water,acetone, ethanol, ethylene glycol, propylene glycol, butane-1,3-diol,isopropyl myristate, isopropyl palmitate, mineral oil, and mixturesthereof to form solutions, emulsions or gels, which are non-toxic andpharmaceutically acceptable. Moisturizers or humectants can also beadded to pharmaceutical compositions and dosage forms if desired.Examples of such additional ingredients are well known in the art. See,e.g., Remington's Pharmaceutical Sciences, 16^(th) and 18^(th) eds.,Mack Publishing, Easton Pa. (1980 & 1990).

The pH of a pharmaceutical composition or dosage form may also beadjusted to improve delivery of one or more active ingredients.Similarly, the polarity of a solvent carrier, its ionic strength, ortonicity can be adjusted to improve delivery. Compounds such asstearates can also be added to pharmaceutical compositions or dosageforms to advantageously alter the hydrophilicity or lipophilicity of oneor more active ingredients so as to improve delivery. In this regard,stearates can serve as a lipid vehicle for the formulation, as anemulsifying agent or surfactant, and as a delivery-enhancing orpenetration-enhancing agent. Different salts, hydrates or solvates ofthe active ingredients can be used to further adjust the properties ofthe resulting composition.

4.4.5 Kits

Typically, active ingredients of the invention are preferably notadministered to a patient at the same time or by the same route ofadministration. This invention therefore encompasses kits which, whenused by the medical practitioner, can simplify the administration ofappropriate amounts of active ingredients to a patient.

A typical kit of the invention comprises a dosage form of a compound ofthis invention. Kits encompassed by this invention can further compriseadditional active ingredients such as oblimersen (Genasense®),melphalan, G-CSF, GM-CSF, EPO, topotecan, dacarbazine, irinotecan,taxotere, WFN, COX-2 inhibitor, pentoxifylline, ciprofloxacin,dexamethasone, IL2, IL8, IL18, Ara-C, vinorelbine, isotretinoin, 13cis-retinoic acid, or a pharmacologically active mutant or derivativethereof, or a combination thereof. Examples of the additional activeingredients include, but are not limited to, those disclosed herein(see, e.g., section 5.2).

Kits of the invention can further comprise devices that are used toadminister the active ingredients. Examples of such devices include, butare not limited to, syringes, drip bags, patches, and inhalers.

Kits of the invention can further comprise cells or blood fortransplantation as well as pharmaceutically acceptable vehicles that canbe used to administer one or more active ingredients. For example, if anactive ingredient is provided in a solid form that must be reconstitutedfor parenteral administration, the kit can comprise a sealed containerof a suitable vehicle in which the active ingredient can be dissolved toform a particulate-free sterile solution that is suitable for parenteraladministration. Examples of pharmaceutically acceptable vehiclesinclude, but are not limited to: Water for Injection USP; aqueousvehicles such as, but not limited to, Sodium Chloride Injection,Ringer's Injection, Dextrose Injection, Dextrose and Sodium ChlorideInjection, and Lactated Ringer's Injection; water-miscible vehicles suchas, but not limited to, ethyl alcohol, polyethylene glycol, andpolypropylene glycol; and non-aqueous vehicles such as, but not limitedto, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate,isopropyl myristate, and benzyl benzoate.

5. EXAMPLES

Certain embodiments of the invention are illustrated by the followingnon-limiting examples.

5.1 2-(4-Amino-1-oxo-1,3-dihydro-isoindol-2-yl)-4-cabamoyl-butyric acid

Step 1: To a suspension of methyl-2-bromomethyl-3-nitrobenzoate (10.6 g,39 mmol) and L-glutamine t-butyl ester hydrochloride (9.2 g, 39 mmol) inTHF (92 mL), was added TEA (12 mL) with stirring. The resulting mixturewas refluxed for 12 hours and then cooled to ambient temperature. Themixture was concentrated to dryness, charged with water (100 mL) andMTBE (100 mL). The slurry was stirred at ambient temperature for 30minutes. The solid was collected by vacuum filtration, washed with 0.2 Naqueous HCl (30 mL), DI water (30 mL×2), MTBE-(30 mL×3), and dried at60° C. in vacuo overnight, giving 9.37 g (67% yield) of4-carbamoyl-2-(4-notro-1-oxo-1,3-dihydro-isoindo-2-yl)-butyric acidt-butyl ester as a white powder. ¹H NMR (DMSO-d₆): 8.48 (d, 1H), 8.17(d, 1H), 7.84 (t, 1H), 7.24 (s, 1H), 6.78 (s, 1H), 4.94 (s, 2H),4.75-4.81 (m, 1H), 2.10-2.33 (m, 4H), 1.41 (s, 9H).

Step 2: TFA (47 mL) was added to a stirred slurry of4-cabamoyl-2-(4-nitro-1-oxo-1,3-dihydro-isoindo-2-yl)-butyric acidt-butylester (9.4 g, 26 mmol) in DCM (47 mL) at −15° C. over 55 minutes.The mixture was stirred at −15° C. for another 5 minutes after theaddition of TFA. It was then allowed to warm to ambient temperature andstirring was continued at ambient temperature for an additional 5 hours.The reaction solution was concentrated in vacuo, stirred with 95 mL ofethyl acetate at ambient temperature for 16 hours. The solid wascollected by vacuum filtration, washed with EtOAc (10 mL×2), and driedin vacuo at 60° C. for 15 hours, affording 7.60 g (95% yield) of a paletan powder of4-carbamoyl-2-(4-nitro-1-oxo-1,3-dihydro-isoindo-2-yl)-butyric acid. ¹HNMR (DMSO-d₆): 13.14 (brs, 1H), 8.47 (d, 1H), 8.17 (d, 1H), 7.84 (t,1H), 7.22 (s, 1H), 6.75 (s, 1H), 4.94 (s, 2H), 4.78-4.84 (m, 1H),2.11-2.37 (m, 4H).

Step 3: 4-Carbamoyl-2-(4-nitro-1-oxo-1,3-isoindo-2-yl)-butyric acid (9.8g, 32 mmol), 10% Pd/C (1 g), methanol (150 mL), and DI water (50 mL)were combined and shaken under 45-50 psi H₂ for 6 hours at ambienttemperature. The reaction slurry was filtered through a celite bed andthe celite bed was washed with 80 mL of methanol. The methanol wash andthe filtrate were combined and concentrated in vacuo to dryness. Theresidue was then dissolved in DI water (200 mL), frozen in a dry-icebath, and lyophilized for one week to provide 8.30 g of an off-whitesolid 2-(4-amino-1-oxo-1,3-dihydro-isoindol-2-yl)-4-carbamoyl-butyricacid in 90% yield: mp 158-160° C.; ¹H NMR DMSO-d₆): 7.14-7.28 (m, 2H),6.76-6.90 (m, 3H), 5.51 (brs, 2H), 4.71-4.77 (m, 1H), 4.16-4.32 (m, 2H),1.90-2.32 (m, 4H) (showed 0.1 mol of MeOH and 0.4 mol H₂O); ¹³C NMR(DMSO-d₆): 173.19, 172.55, 169.06, 143.64, 132.41, 128.81, 125.74,116.32, 110.44, 53.22, 48.65 (MeOH), 45.26, 31.60, 24.96. Analy.calculated for C₁₃H₁₃N₃O₄-0.4H₂O/0.1MeOH: 54.69% C, 5.68% H, 14.61% N,2.5% H₂O. Found: 55.05% C, 5.42% H, 14.39% N, 3.6% H₂O.

5.2 4-(4-Amino-1-oxo-1,3-dihydro-isoindol-2-yl)-4-carbamoyl-butyric acid

Step 1: To a suspension of methyl-2-bromomethyl-3-nitrobenxoate (5.8 g,21 mmol) and L-isoglutamine t-butyl ester hydrochloride (5 g, 21 mmol)in THF (50 mL), was added TEA (6.5 mL) with stirring. The resultingmixture was refluxed for 24 hours and then cooled to ambienttemperature. The mixture was concentrated to dryness, charged with water(75 mL) and MTBE (75 mL). The slurry was then stirred at ambienttemperature for 2 hours. The solid was collected by vacuum filtration,washed with 0.2 N aqueous HCl (15 mL), DI water (20 mL×7 to neutral pH),and MTBE (20 mL). The wet cake was further reslurried at ambienttemperature for 16.5 hours in a mixture of solvent containing MTBE (100mL) and DI water (50 mL). The solid was collected by vacuum filtration,washed with MTBE (65 mL), and dried at 60° C. in vacuo for 15 hours,giving 5.30 g (70% yield) of4-carbamoyl-4-(4-nitro-1-oxo-1,3-dihydro-isoindo-2-yl)-butyric acidt-butyl ester as a pale tan powder. mp 174-176° C.; ¹H NMR (DMSO-d₆):8.45 (d, 1H), 8.16 (d, 1H), 7.82 (t, 1H), 7.64 (s, 1H), 7.27 (s, 1H),4.98 (dd, 2H), 4.78 (dd, 1H), 2.00-2.25 (m, 4H), 1.33 (s, 9H); ¹³C NMR(DMSO-d₆): 171.49, 171.37, 165.81, 143.27, 137.80, 135.00, 129.91,129.56, 126.73, 79.80, 53.75, 48.17, 31.65, 27.65, 24.83 ppm. Analy.calculated for C₁₇H₃₁N₃O₆: 56.19% C, 5.83% H, 11.56% N. Found: 55.89% C,5.53% H, 11.26% N.

Step 2: TFA (26 mL) was added to a stirred slurry of4-carbamoyl-4-(4-nitro-1-oxo-1,3-dihydro-isoindo-2-yl)-butyric acidt-butyl ester (5.2 g, 14 mmol) in DCM (26 mL) at −15° C. over 25minutes. The mixture was stirred at −15° C. for another 5 minutes afterthe addition of TFA. It was then allowed to warm to ambient temperatureand stirring was continued at ambient temperature for an additional 4hours. The reaction solution was concentrated in vacuo, stirred with 52mL of ethyl acetate at ambient temperature for 15 hours. The solid wascollected by vacuum filtration, washed with EtOAc twice (14+21 mL), andreslurried in acetone (20 mL) at ambient temperature for 2 hours. Thesuspension was filtered and the solid was washed with acetone (27 mL×2).The solid was dried in vacuo at 60° C. for 2 hours, affording 4.17 g(95% yield) of4-carbamoyl-4-(4-nitro-1-oxo-1,3-dihydro-isoindo-2-yl)-butyric acid as apale tan powder. mp 219-221° C.; ¹H NMR (DMSO-d₆): 12.14 (brs, 1H), 8.46(d, 1H), 8.16 (d, 1H), 7.82 (t, 1H), 7.67 (s, 1H), 7.29 (s, 1H), 5.00(dd, 2H), 4.77-4.82 (m, 1H), 1.99-2.26 (m, 4H); ¹³C NMR (DMSO-d₆):173.63, 171.65, 165.89, 143.31, 137.85, 135.00, 129.92, 129.59, 126.76,53.78, 48.19, 30.58, 24.97. Analy. calculated for C₁₃H₁₃N₃O₆.0.2H₂O:50.23% C, 4.34% H, 13.52% N, 1.2% H₂O. Found: 50.23% C, 4.06% H, 13.23%N, 1.2% H₂O.

Step 3: 4-Carbamoyl-4-(4-nitro-1-oxo-1,3-dihydro-isoindo-2-yl)-butyricacid (8.8 g, 29 mmol), 10% Pd/C (0.88 g), methanol (132 mL), and DIwater (44 mL) were combined and shaken under 45-50 psi H₂ for 6 hours atambient temperature. The reaction slurry was filtered through a celitebed and the celite bed was washed with 75 mL of methanol. The methanolwash and the filtrate were combined and concentrated in vacuo todryness. DI water (100 mL) was added to the residue, and the mixture wasagain concentrated to dryness. The residual oily material was thendissolved in DI water (200 mL), frozen in a dry-ice bath, andlyophilized for 8 days to provide 7.30 g of4-(4-amino-1-oxo-1,3-dihydro-isoindol-2-yl)-4-carbamoyl-butyric acid asan off-white solid in 92% yield. mp 334-336° C.; ¹H NMR (DMSO-d₆): 12.07(brs, 1H), 7.54 (s, 1H), 6.76-7.20 (m, 4H), 5.42 (brs, 2H), 4.71-4.75(m, 1H), 4.32 (dd, 2H), 1.90-2.22 (m, 4H); ¹³C NMR (DMSO-d₆): 173.62,172.16, 168.84, 143.39, 132.48, 128.68, 126.06, 116.28, 110.53, 53.26,45.31, 30.69, 25.09. Analy. calculated for C₁₃H₁₅N₃O₄.0.2H₂O: 55.59% C,5.53% H, 14.96% N, 1.3% H₂O. Found: 55.69% C, 5.30% H, 14.83% N, 1.4%H₂O.

5.34-Carbamoyl-4-[4-[(furan-2-yl-methyl)-amino]-1,3-dioxo-1,3-dihydro-isoindol-2-yl]-butyricacid 5.3.1 3-[(Furan-2-ylmethyl)-amino]-phthalic acid dimethyl ester

To a stirred solution of 3-amino-phthalic acid dimethyl ester (8.23 g,39.32 mmol) in methylene chloride (200 ml) under a nitrogen atmosphere,2-furaldehyde (8.14 ml, 98.30 mmol) and acetic acid (13.57 ml, 235.92mmol) were added. The mixture was stirred for 5 minutes, followed byaddition of sodium triacetoxyborohydride (25 g, 117.96 mmol). Thereaction was stirred overnight, washed with water (2×200 ml), saturatedaqueous sodium bicarbonate (2×200 ml), and brine (200 ml), and driedover MgSO₄. The solvent was evaporated in vacuo to give a brown oil(12.42 g), which was used directly without purification.

5.3.2 3-[(Furan-2-yl-methyl)-amino]-phtalic acid

To a stirred solution of crude 3-[(furan-2-yl-methyl)-amino]-phthalicacid dimethyl ester in methanol (100 ml), was added 5N potassiumhydroxide (79 ml). The mixture was stirred at room temperatureovernight. The solvent was evaporated in vacuo and the residue dissolvedin water (50 ml). The water was washed with diethyl ether (2×100 ml).The aqueous portion was cooled in an ice bath and the pH was adjusted to2-3 by dropwise addition of concentrated hydrochloric acid. The aqueoussolution was then extracted into ethyl acetate (3×100 ml). The combinedethyl acetate extracts were washed with brine (150 ml), and dried overMgSO₄. The solvent was evaporated in vacuo, and the residue, whichcontained a mixture of diacid and monomethyl esters, was used withoutfurther purification.

5.3.3 4-Carbamoyl-4-{4-[(furan-2-yl-methylamino]-1,3-dioxo1,3-dihydro-isoindol-2-yl}-butyric acid

To a stirred solution of 3-[(furan-2-yl-,ethyl)-amino]-phthalic acid(0.5 g, 1.93 mmol) in pyridine (20 ml), was added4-amino-4-carbamoyl-butyric acid (0.31 g, 2.12 mmol). The reactionmixture was heated to reflux overnight. The solvent was evaporated invacuo. The resulting residue was dissolved in saturated sodiumbicarbonate (50 ml) and washed with ethyl acetate (2×50 ml). The aqueousportion was cooled in an ice bath, and the pH was adjusted to 2-3 bydropwise addition of concentrated hydrochloric acid. The aqueoussolution was then extracted into ethyl acetate (3×50 ml). The combinedethyl acetate extracts were washed with brine (100 ml), and dried overMgSO₄. The solvent was evaporated in vacuo to give an oil. The oil waspurified by flash column chromatography (60% ethyl acetate/39% hexane/1%formic acid) to give an oil, which was triturated in diethyl ether (30ml). The resulting yellow solid was filtered and dried (0.14 g, 20%): mp120-122° C.; ¹H NMR (DMSO-d₆) δ 12.09 (s, 1H), 7.59-7.62 (m, 3H), 7.15(d, J=8.25 Hz, 2H), 7.03-6.93 (m, 2H), 6.41-6.36 (m, 2H), 4.56-4.50 (m,3H), 2.40-2.17 (m, 4H); ¹³C NMR (DMSO-d₆) δ 173.57, 170.04, 169.36,167.71, 152.07, 145.57, 142.41, 135.60, 132.55, 116.99, 110.70, 110.46,110.42, 107.41, 51.73, 30.66, 23.52; Analy. Calculated for C₁₈H₁₇N₃O₆:C, 57.25; H, 4.72; N, 11.13 (+0.35H₂O). Found: C, 57.50; H, 4.59; N,10.93.

5.44-Carbamoyl-2-{4-[(furan-2-yl-methyl)-amino]-1,3-dioxo-1,3-dihydro-isoindol-2-yl}-butyricacid

2-Amino-4-carbamoyl-butyric acid (0.31 g, 2.11 mmol) was treated usingthe procedures substantially the same as those described in Section 5.3,above, for the synthesis of4-carbamoyl-2-{4-[(furan-2-yl-methyl)-amino]-1,3-dioxo-1,3-dihydro-isoindol-2-yl}-butyricacid. The residue was purified by flash column chromatography (66% ethylacetate/33% hexane/1% formic acid) to give an oil (0.44 g). Furtherpurification by preparative reverse phase HPLC (38% acetonitrile/62%water, isocratic) provided an oil, which was triturated in diethyl ether(30 ml). The resulting yellow solid was filtered and dried (0.31 g,44%): mp 114-116° C.; ¹H NMR (DMSO-d₆) δ 13.14 (s, 1H), 7.59-7.54 (m,2H), 7.19-7.16 (m, 2H), 7.05-6.97 (m, 2H), 6.70 (s, 1H), 6.41-6.36 (m,2H), 4.64 (dd, J=4.49 Hz and J=10.49 Hz, 1H), 4.55 (d, J=6.01 Hz, 2H),2.42-2.17 (m, 2H), 2.14-2.02 (m, 2H); ¹³C NMR (DMSO-d₆) δ 173.03,170.61, 169.06, 167.48, 151.99, 145.72, 142.44, 135.94, 132.13, 117.39,110.89, 110.43, 109.79, 107.43, 50.93, 31.35, 24.02, 15.14; Analy.Calculated for C₁₉H₁₇N₃O₆: C, 57.38; H, 4.71; N, 11.15 (+0.30H₂O).Found: C, 57.00; H, 4.85; N 10.83.

5.52-{4-[(Furan-2-yl-methyl)-amino]-1,3-dioxo-1,3-dihydro-isoindol-2-yl}4-phenylcarbamoyl-butyricacid

2-Amino-4-phenylcarbamoyl-butyric acid (0.49 g, 2.20 mmol) was treatedaccording to the procedures substantially the same as those described inSection 5.3, above, for the synthesis of2-{4-[(furan-2-yl-methyl)-amino]-1,3-dioxo-1,3-dihydro-isoindol-2-yl}-4-phenylcarbamoyl-butyricacid. The residue was purified by flash column chromatography (45% ethylacetate/54% hexane/1% formic acid) to provide a yellow solid (0.48 g,53.6%): mp 146-148° C.; ¹H NMR (DMSO-d₆) δ 13.13 (s, 1H), 9.81 (s, 1H),7.59-7.38 (m, 4H), 7.26-7.14 (m, 3H), 7.04-6.96 (m, 3H), 6.40-6.34 (m,2H), 4.71 (dd, J=3.80 Hz and J=9.50 Hz, 1H), 4.54 (d, J=5.95 Hz, 2H),2.46-2.22 (m, 4H); ¹³C NMR (DMSO-d₆) δ 170.51, 169.95, 169.05, 167.46,151.99, 145.73, 142.42, 139.07, 135.93, 132.12, 128.50, 122.88, 119.01,117.39, 110.91, 110.43, 109.80, 107.39, 50.78, 32.75, 23.97; Analy.Calculated for C₂₄H₂₁N₃O₆: C, 63.63; H, 4.81; N, 9.28 (+0.31H₂O). Found:C, 63.38; H, 4.84; N, 9.20.

5.62-[4-[(Furan-2-yl-methyl)-amino]-1,3-dioxo-1,3-dihydro-isoindol-2-yl]-pentanedioicacid

2-Amino-pentanedioic acid (0.32 g, 2.18 mmol) was treated usingprocedures substantially the same as those described in Section 5.3,above, for the synthesis of2-{4-[(furan-2-yl-methyl)-amino]-1,3-dioxo-1,3-dihydro-isoindol-2-yl}-pentanedioicacid. The residue was purified by flash column chromatography (42.5%ethyl acetate/56.5% hexane/1% formic acid) to provide a yellow solid(0.54 g, 73%): mp 150-152° C.; ¹H NMR (DMSO-d₆) δ 12.62 (s, 2H), 9.81(s, 1H), 7.60-7.54 (m, 2H), 7.17 (d, J=8.58 Hz, 1H), 7.05-6.98 (m, 2H),6.41-6.36 (m, 2H), 4.72 (dd, J=4.10 Hz and J=9.80 Hz, 1H), 4.56 (d,J=6.05 Hz, 2H), 2.38-2.20 (m, 4H); ¹³C NMR (DMSO-d₆) δ 173.59, 170.47,169.04, 167.46, 151.98, 145.74, 142.42, 135.94, 132.10, 117.42, 110.93,110.42, 109.77, 107.40, 95.55, 50.55, 30.29, 23.69; Analy. Calculatedfor C₁₈H₁₆N₂O₇: C, 57.40; H, 4.40; N, 7.40 (+0.17H₂O+0.02 EtOAc+0.02CH₂Cl₂). Found: C, 57.01; H, 4.18; N, 7.27.

5.7 Cyclopropanecarboxylic acid{2-[1-(3-ethoxy-4-methoxy-phenyl)-2-[1,3,4]oxadiazol-2-yl-ethyl]-3-oxo-2,3-dihydro-1H-isoindol-4-yl}-amide

Cyclopropanecarboxylic acid{2-[1-(3-ethoxy-4-methoxy-phenyl)-2-[1,3,4]oxadiazol-2-yl-ethyl]-3-oxo-2,3-dihydro-1H-isoindol-4-yl}-amidewas prepared by reacting3-[7-(cyclopropanecarbonyl-amino)-1-oxo-1,3-dihydro-isoindol-2-yl]-3-(3-ethoxy-4-methoxy-phenyl)-propionicacid (400 mg, 0.9 mmol), CDI (160 mg, 1.0 mmol) and formic hydrazide(0.09 g, 1.5 mmol) in THF (5 mL). The crude was then reacted withphosphorus oxychloride (0.18 mL, 1.9 mmol) in acetonitrile (15 mL). Theproduct was obtained as a white solid (60 mg, 15% yield): mp 181-183°C.; ¹H NMR (CDCl₃) δ 0.86-0.92 (m, 2H, cyclopropyl CH₂), 1.06-1.11 (m,2H, cyclopropyl CH₂), 1.45 (t, J=7.5 Hz, 3H, CH₃), 1.58-1.68 (m, 1H,cyclopropyl CH), 3.66 (dd, J=5, 15 Hz, 1H, CHH), 3.85 (dd, J=10, 15 Hz,1H, CHH), 3.87 (s, 3H, CH₃), 4.03-4.11 (m, 3H, OCH₂CH₃+CHH), 4.43 (d,J=17 Hz, 1H, CHH), 5.86-5.91 (m, 1H, NCH), 6.85-6.90 (m, 2H, Ar),6.96-7.01 (m, 2H, Ar), 7.43 (t, J=7.5 Hz, 1H, Ar), 8.35 (s, 1H, Ar),8.41 (d, J=7.5 Hz, 1H, Ar), 10.41 (s, 1H, NH); ¹³C NMR (CDCl₃) , 8.27,8.29, 14.73, 16.16, 28.04, 46.36, 52.26, 55.98, 64.64, 111.53, 112.36,116.80, 116.96, 117.80, 119.16, 129.54, 133.43, 138.06, 141.25, 148.84,149.64, 153.34, 163.79, 169.50, 172.66. Analy. calculated forC₂₅H₂₅N₄O₅+0.1H₂O: C, 64.47; H, 5.69; N, 12.07. Found: C, 64.50; H,5.70; N, 11.69.

5.8 Inhibition of MM Cell Proliferation

The ability of a compound of this invention to effect the proliferationof multiple myeloma (MM) cell lines is investigated in an in vitrostudy. Uptake [³H]-thymidine by different MM cell lines (MM. IS, HsSultan, U266 and RPMI-8226) is measured as an indicator of cellproliferation. Cells are incubated in the presence of compound for 48hours; [³H]-thymidine is included for the last 8 hours of the incubationperiod.

5.9 In Vivo LPS-Induced TNF-α Production Assay

Male CD rats procured from Charles River Laboratories at seven weeks ofage are allowed to acclimate for one week prior to use. A lateral tailvein is cannulated percutaneously with a 22-gage over-the-needlecatheter under brief isoflurane anesthesia. Rats are administered acompound of this invention either by intravenous injection via the tailvein catheter or oral gavage 15 to 180 min prior to injection of 0.05mg/kg LPS (E. Coli 055:B5). Catheters are flushed with 2.5 mL/kg ofnormal injectable saline. Blood is collected via cardiac puncture 90minutes after LPS challenge. Plasma is prepared using lithium heparinseparation tubes and frozen at −80° C. until analyzed. TNF-α levels aredetermined using a rat specific TNF-α ELISA kit (Busywork). The ED₅₀values are calculated as the dose of the compound of this invention atwhich the TNF-α production is reduced to 50% of the control value.

5.10 Cycling Therapy in Patients

In a specific embodiment, a compound of this invention are cyclicallyadministered to patients with cancer. Cycling therapy involves theadministration of a first agent for a period of time, followed by a restfor a period of time and repeating this sequential administration.Cycling therapy can reduce the development of resistance to one or moreof the therapies, avoid or reduce the side effects of one of thetherapies, and/or improves the efficacy of the treatment.

In a specific embodiment, prophylactic or therapeutic agents areadministered in a cycle of about four to six weeks, about once or twiceevery day. One cycle can comprise the administration of a therapeutic onprophylactic agent for three to four weeks and at least one week or twoweeks of rest. The number of cycles administered is from about one toabout 24 cycles, more typically from about two to about 16 cycles, andmore typically from about four to about eight cycles.

5.11 Clinical Studies in Patients with Relapsed Multiple Myeloma

Patients with relapsed and refractory Dune-Salmon stage III multiplemyeloma, who have either failed at least three previous regimens orpresented with poor performance status, neutropenia or thrombocytopenia,are treated with up to four cycles of combination melphalan (50 mgintravenously), a compound of this invention (about 1 to 5,000 mg orallydaily), and dexamethasone (40 mg/day orally on days 1 to 4) every fourto six weeks. Maintenance treatment consisting of daily a compound ofthis invention and monthly dexamethasone are continued until the diseaseprogression. The therapy comprising the administration of a compound ofthis invention in combination with melphalan and dexamethasone is highlyactive and generally tolerated in heavily pretreated multiple myelomapatients whose prognosis is otherwise poor.

The embodiments of the invention described above are intended to bemerely exemplary, and those skilled in the art will recognize, or willbe able to ascertain using no more than routine experimentation,numerous equivalents of specific compounds, materials, and procedures.All such equivalents are considered to be within the scope of theinvention and are encompassed by the appended claims.

All of the patents, patent publications and references cited herein areincorporated in their entirety by reference. However, citation of suchreferences do not constitute an admission that such references are priorart. This invention can be better illustrated by the following claims.

1. A compound of formula:

or a pharmaceutically acceptable salt or stereoisomer thereof, whereinR¹ and R², independently of the other, is hydroxyl or NH—Z; R³ ishydrogen; Z is hydrogen or aryl; and n is 0, 1 or
 2. 2. A pharmaceuticalcomposition comprising a compound of claim 1.